論文 - 多胡　めぐみ

The critical role of the phosphorylation of STAT3 at Y705 in ALCL-associated NPM-ALK-induced transforming activity

Lin X., Korai A., Nakazawa Y., Tago K., Funakoshi-Tago M.

Cellular Signalling 136 2025年12月

ISSN  08986568

　概要を見る

The fusion protein nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) drives oncogenesis in anaplastic large cell lymphoma (ALCL) by activating signal transducer and activator of transcription 3 (STAT3). NPM-ALK requires its kinase activity to induce STAT3 phosphorylation at tyrosine 705 (Y705) and promotes serine 727 (S727) phosphorylation via JNK activation. However, the role of these modifications in NPM-ALK-driven cellular transformation remains unclear. We herein utilized murine Ba/F3 cells expressing NPM-ALK to investigate the impact of these modifications. STAT3 knockdown, followed by reconstitution with wild-type STAT3 or its mutants (Y705F and S727A) revealed that Y705 phosphorylation was essential for NPM-ALK-mediated transformation. STAT3 knockdown suppressed the expression of NPM-ALK-induced STAT3 target genes (c-Myc, Pim, IL-6, and SOCS3) as well as cell proliferation, tumor formation, and spleen, liver, and lymph node enlargement. These effects were restored upon reconstitution with wild-type STAT3 or the S727A mutant, but not the Y705F mutant, confirming the essential role of Y705 phosphorylation in these biological processes. Additionally, wild-type and mutant STAT3 proteins exhibited differential stability, with Y705F being less stable and S727A being more stable. Lysosomal inhibition by bafilomycin A1 increased the expression of wild-type STAT3 and the Y705F mutant, but had no effect on the S727A mutant. Cycloheximide chase assays further confirmed that S727 phosphorylation regulated STAT3 degradation via the lysosomal pathway. These results identify a novel NPM-ALK-induced oncogenic mechanism mediated by STAT3 phosphorylation, highlighting potential therapeutic targets for ALCL.

• Access to Document (DOI)

TRPV1 attenuates epithelial-mesenchymal transition via calpain-protein tyrosine phosphatase pathway in lens epithelial cells

Nakazawa Y., Nishizawa F., Kawata S., Sugiyama Y., Nagai N., Yamamoto N., Funakoshi-Tago M.

Experimental Eye Research 258 2025年09月

ISSN  00144835

　概要を見る

TRPV1, which is widely expressed throughout the body, is a non-selective cation channel activated by capsaicin. We previously reported that TRPV1 activation suppressed TGFβ2-induced epithelial-mesenchymal transition (EMT) by inhibiting Epidermal Growth Factor Receptor (EGFR) phosphorylation in lens epithelial cells (Sugiyama et al., 2021). However, the detailed molecular mechanism remains unclear. In this study, we focused on the calpain–protein tyrosine phosphatase (PTP) pathway to elucidate the detailed mechanism underlying TRPV1-induced EMT suppression. Calpain and PTP inhibitors mitigated the suppressive effect of capsaicin on TGFβ2-induced EMT in vitro and ex vivo. Finally, we shown that CalpainS1 and PTPN9 overexpression abrogated the effect of capsaicin on EMT in lens epithelial cells. Our findings indicate that calpain and PTP proteins are good candidates for preventing EMT after cataract surgery.

• Access to Document (DOI)

Development of Keap1-Nrf2 Protein–Protein Interaction Inhibitor Activating Intracellular Nrf2 Based on the Naphthalene-2-acetamide Scaffold, and its Anti-Inflammatory Effects

Yasuda D., Toyoshima K., Kojima K., Ishida H., Kaitoh K., Imamura R., Kanamitsu K., Kojima H., Funakoshi-Tago M., Osawa M., Ohe T., Hirano T.

Chemmedchem 2025年

ISSN  18607179

　概要を見る

Nuclear factor erythroid 2-related factor 2 (Nrf2) and Kelch-like ECH-associated protein 1 (Keap1) axis is an attractive therapeutic target for various intractable diseases. Although protein–protein interaction inhibitors against Keap1-Nrf2 have been developed over the past decade, more structural expansion is needed to improve efficacy. In this article, several candidate compounds are designed and synthesized as novel Nrf2 activators and their intracellular Nrf2-activating effects are evaluated. Among the synthesized compounds, a novel naphthalene-1,4-(4-ethoxybenzensulfonamide) bearing a tertiary acetamide side chain at the 2-position strongly activated intracellular Nrf2. Particularly, the pyrrolidine-type acetamide compound showed the strongest intracellular Nrf2 activation. X-ray cocrystallography revealed that this compound can bind to the DC domain of Keap1. Additionally, the pyrrolidine-type acetamide compound induced the mRNA expression of the representative Nrf2 target genes heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1. Moreover, the compound exhibited anti-inflammatory effects in a lipopolysaccharide-stimulated macrophage cell line. Conclusively, these results suggest that the pyrrolidine-type naphthalene-2-acetamide is a promising compound for the development of Nrf2 activators that can be applied to treat inflammatory diseases.

• Access to Document (DOI)

High ambient temperature may induce presbyopia via TRPV1 activation

Nakazawa Y., Kuno Y., Shimada H., Nagai N., Hiramatsu N., Takeda S., Yamamoto N., Funakoshi-Tago M., Sasaki H.

Medical Molecular Morphology 57 （ 4 ） 268 - 276 2024年12月

ISSN  18601480

　概要を見る

The prevalence of presbyopia and nuclear cataracts (NUC) is reported to be higher in tropical areas than that in other regions, suggesting a potential influence of high temperatures on lens health. Transient receptor potential vanilloid (TRPV) channels play a crucial role in detecting ambient temperatures across various species, with TRPV1 and TRPV4 expressed in lens epithelial cells. In this study, we investigated whether ambient temperatures affect TRPV1 and TRPV4 activity in the lens, potentially contributing to the development of presbyopia and NUC. We conducted experiments using cultured human lens epithelial cell lines under different temperature conditions. Our results revealed that the mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) and p38 pathways, downstream molecules of TRPV1, were activated, while Src family kinase, a downstream molecule of TRPV4, was inhibited at 37.5 °C culture compared to 35.0 °C. Confocal microscope images demonstrated higher expression of TRPV1 in 3D-structured cells under high-temperature culture conditions. Additionally, in organ culture lenses, higher elasticity was observed at elevated temperatures compared to that at lower temperatures. These results suggest that high ambient temperatures may induce lens sclerosis via TRPV1 activation, potentially contributing to the development of presbyopia and NUC.

• Access to Document (DOI)

FL118 Is a Potent Therapeutic Agent against Chronic Myeloid Leukemia Resistant to BCR-ABL Inhibitors through Targeting RNA Helicase DDX5

Takeda K., Ohta S., Nagao M., Kobayashi E., Tago K., Funakoshi-Tago M.

International Journal of Molecular Sciences 25 （ 7 ）  2024年04月

ISSN  16616596

　概要を見る

Chronic myeloid leukemia (CML) is induced by the expression of the fused tyrosine kinase BCR-ABL, which is caused by a chromosomal translocation. BCR-ABL inhibitors have been used to treat CML; however, the acquisition of resistance by CML cells during treatment is a serious issue. We herein demonstrated that BCR-ABL induced the expression of the RNA helicase DDX5 in K562 cells derived from CML patients in a manner that was dependent on its kinase activity, which resulted in cell proliferation and survival. The knockout of DDX5 decreased the expression of BIRC5 (survivin) and activated caspase 3, leading to apoptosis in K562 cells. Similar results were obtained in cells treated with FL118, an inhibitor of DDX5 and a derivative compound of camptothecin (CPT). Furthermore, FL118 potently induced apoptosis not only in Ba/F3 cells expressing BCR-ABL, but also in those expressing the BCR-ABL T315I mutant, which is resistant to BCR-ABL inhibitors. Collectively, these results revealed that DDX5 is a critical therapeutic target in CML and that FL118 is an effective candidate compound for the treatment of BCR-ABL inhibitor-resistant CML.

• Access to Document (DOI)

The acetylation of STAT3 at K685 attenuates NPM-ALK-induced tumorigenesis

Korai A., Lin X., Tago K., Funakoshi-Tago M.

Cellular Signalling 114 2024年02月

ISSN  08986568

　概要を見る

Nucleophosmin-anaplastic lymphoma kinase (NPM-ALK), a fusion protein generated by a chromosomal translocation, is a causative gene product of anaplastic large cell lymphoma (ALCL). It induces cell proliferation and tumorigenesis by activating the transcription factor, signal transducer and activator of transcription factor 3 (STAT3). We herein demonstrated that STAT3 underwent acetylation at K685 in a manner that was dependent on the kinase activity of NPM-ALK. To investigate the role of STAT3 acetylation in NPM-ALK-induced oncogenesis, we generated Ba/F3 cells expressing NPM-ALK in which STAT3 was silenced by shRNA, named STAT3-KD cells, and then reconstituted wild-type STAT3 or the STAT3 K685R mutant into these cells. The phosphorylation level of the K685R mutant at Y705 and S727 was significantly higher than that of wild-type STAT3 in STAT3-KD cells. The expression of STAT3 target genes, such as IL-6, Pim1, Pim2, and Socs3, was more strongly induced by the reconstitution of the K685R mutant than wild-type STAT3. In addition, the proliferative ability of STAT3-KD cells reconstituted with the K685R mutant was slightly higher than that of STAT3-KD cells reconstituted with wild-type STAT3. In comparisons with the inoculation of STAT3-KD cells reconstituted with wild-type STAT3, the inoculation of STAT3-KD cells reconstituted with the K685R mutant significantly enhanced tumorigenesis and hepatosplenomegaly in nude mice. Collectively, these results revealed for the first time that the acetylation of STAT3 at K685 attenuated NPM-ALK-induced oncogenesis.

• Access to Document (DOI)

Suppression of Neuroinflammation by Coffee Component Pyrocatechol via Inhibition of NF-κB in Microglia

Murata T., Tago K., Miyata K., Moriwaki Y., Misawa H., Kobata K., Nakazawa Y., Tamura H., Funakoshi-Tago M.

International Journal of Molecular Sciences 25 （ 1 ）  2024年01月

ISSN  16616596

　概要を見る

According to numerous studies, it has been epidemiologically suggested that habitual coffee intake seems to prevent the onset of neurodegenerative diseases. In this study, we hypothesized that coffee consumption suppresses neuroinflammation, which is closely related to the development of neurodegenerative diseases. Using microglial BV-2 cells, we first found that the inflammatory responses induced by lipopolysaccharide (LPS) stimulation was diminished by both coffee and decaffeinated coffee through the inhibition of an inflammation-related transcription factor, nuclear factor-κB (NF-κB). Pyrocatechol, a component of roasted coffee produced by the thermal decomposition of chlorogenic acid, also exhibited anti-inflammatory activity by inhibiting the LPS-induced activation of NF-κB. Finally, in an inflammation model using mice injected with LPS into the cerebrum, we observed that intake of pyrocatechol as well as coffee decoctions drastically suppressed the accumulation of microglia and the expression of interleukin-6 (IL-6), tumor necrosis factor α (TNFα), CCL2, and CXCL1 in the inflammatory brain. These observations strongly encourage us to hypothesize that the anti-inflammatory activity of pyrocatechol as well as coffee decoction would be useful for the suppression of neurodegeneration and the prevention of the onsets of Alzheimer’s (AD) and Perkinson’s diseases (PD).

• Access to Document (DOI)

Piezo1 channel causes lens sclerosis via transglutaminase 2 activation

Doki Y., Nakazawa Y., Sukegawa M., Petrova R.S., Ishida Y., Endo S., Nagai N., Yamamoto N., Funakoshi-Tago M., Donaldson P.J.

Experimental Eye Research 237 2023年12月

ISSN  00144835

　概要を見る

Presbyopia is caused by age-related lenticular hardening, resulting in near vision loss, and it occurs in almost every individual aged ≥50 years. The lens experiences mechanical pressure during for focal adjustment to change its thickness. As lenticular stiffening results in incomplete thickness changes, near vision is reduced, which is known as presbyopia. Piezo1 is a mechanosensitive channel that constantly senses pressure changes during the regulation of visual acuity, and changes in Piezo1 channel activity may contribute to presbyopia. However, no studies have reported on Piezo1 activation or the onset of presbyopia. To elucidate the relevance of Piezo1 activation and cross-linking in the development of presbyopia, we analysed the function of Piezo1 in the lens. The addition of Yoda1, a Piezo1 activator, induced an increase in transglutaminase 2 (TGM2) mRNA expression and activity through the extra-cellular signal-regulated kinase (ERK) 1/2 and c-Jun-NH2-terminal kinase1/2 pathways. In ex vivo lenses, Yoda1 treatment induced γ-crystallin cross-linking via TMG2 activation. Furthermore, Yoda1 eye-drops in mice led to lenticular hardening via TGM2 induction and activation in vivo, suggesting that Yoda1-treated animals could serve as a model for presbyopia. Our findings indicate that this presbyopia-animal model could be useful for screening drugs for lens-stiffening inhibition.

• Access to Document (DOI)

The citrus flavonoid, nobiletin inhibits neuronal inflammation by preventing the activation of NF-κB

Murata T., Ishiwa S., Lin X., Nakazawa Y., Tago K., Funakoshi-Tago M.

Neurochemistry International 171 2023年12月

ISSN  01970186

　概要を見る

Nobiletin (5,6,7,8,3′,4′-hexamethoxyflavone) is one of the flavonoids found in shikuwasa, a popular citrus fruit in Okinawa, Japan. It exerts various pharmacological effects, such as anti-tumor, antioxidant, and anti-inflammatory activities. We herein investigated whether nobiletin attenuated lipopolysaccharide (LPS)-induced inflammatory responses in the murine microglial cell line BV-2 and neuroinflammation in mice induced by an intracerebral injection of LPS. In BV-2 cells, nobiletin significantly inhibited the LPS-induced production of nitric oxide (NO) and prostaglandin E2 (PGE2) by preventing the mRNA expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), respectively. Nobiletin also inhibited the LPS-induced mRNA expression of CCL2, CXCL1, IL-6, and TNFα. Nobiletin markedly attenuated the transcriptional activity of the NF-κB p65 subunit without affecting the degradation of IκBα or the nuclear localization of the NF-κB p65 subunit. Nobiletin also inhibited the LPS-induced activation of JNK, but not ERK or p38, in BV-2 cells. Furthermore, the administration of nobiletin significantly suppressed the accumulation of microglia and induction of the mRNA expression of CCL2, CXCL1, IL-6, and TNFα in the murine brain induced by injecting LPS into the striatum. Collectively, these results suggest the potential of nobiletin as a candidate anti-inflammatory drug for the prevention of neuroinflammation.

• Access to Document (DOI)

Aquaporins contribute to vacuoles formation in Nile grass type II diabetic rats

Aihara K., Nakazawa Y., Takeda S., Hatsusaka N., Onouchi T., Hiramatsu N., Nagata M., Nagai N., Funakoshi-Tago M., Yamamoto N., Sasaki H.

Medical Molecular Morphology 56 （ 4 ） 274 - 287 2023年12月

ISSN  18601480

　概要を見る

Regulation of ion and water microcirculation within the lens is tightly controlled through aquaporin channels and connexin junctions. However, cataracts can occur when the lens becomes cloudy. Various factors can induce cataracts, including diabetes which is a well-known cause. The most common phenotype of diabetic cataracts is a cortical and/or posterior subcapsular opacity. In addition to the three main types and two subtypes of cataracts, a vacuole formation is frequently observed; however, their origin remains unclear. In this study, we focused on the aquaporins and connexins involved in diabetes-induced cataracts and vacuoles in Nile grass type II diabetes. The results showed that the expression of aquaporin 0 and aquaporin 5 increased, and that of connexin 43 decreased in diabetic rat lenses. Additionally, aquaporin 0 and 5 were strongly localized in peripheral of vacuoles, suggesting that aquaporins are involved in vacuoles formation. Transillumination photography revealed large vacuoles at the tip of the Y-suture in the anterior capsule of the diabetic lens, and several small vacuoles were observed in the posterior capsule. Within the vacuoles, cytoplasmic degradation and aggregation of fibrous material were observed. Our findings suggest that aquaporins are potential candidate proteins for preventing vacuole formation.

• Access to Document (DOI)

Hesperetin treatment attenuates glycation of lens proteins and advanced-glycation end products generation

Doki Y., Nakazawa Y., Morishita N., Endo S., Nagai N., Yamamoto N., Tamura H., Funakoshi-Tago M.

Molecular Medicine Reports 27 （ 5 ）  2023年05月

ISSN  17912997

　概要を見る

Advanced glycation end products (AGEs) in lens proteins increase with aging, thus inducing cataracts and/or presbyopia. Hesperetin (Hst), which is an abundant plant flavanone largely derived from citrus species, and its derivatives attenuate cataracts and presbyopia in vivo and in vitro; however, no reports have described its effects on AGE formation in lens proteins. The present study demonstrated that AGEs in lens proteins increase with age in mice. Additionally, it showed that Hst can prevent AGEs and N(ϵ)-carboxymethyl-lysine generation and modification of lens proteins using in vitro in human lens epithelial cell lines and ex vivo in mouse lens organ cultures. Furthermore, treatment with Hst prevented lens hardening and decreased chaperone activity in lens proteins. These results suggested that Hst and its derivatives are good candidates for the prevention of presbyopia and cataracts.

• Access to Document (DOI)

The indispensable role of the RNA helicase DDX5 in tumorigenesis induced by the myeloproliferative neoplasm-associated JAK2V617F mutant

Takeda K., Tago K., Funakoshi-Tago M.

Cellular Signalling （Cellular Signalling）  102 2023年02月

ISSN  08986568

　概要を見る

A point mutation (V617F) in the Janus kinase 2 (JAK2) gene results in the production of disorderly activated tyrosine kinase, which causes myeloproliferative neoplasms (MPN). We herein demonstrated that the RNA helicase DDX5 was highly expressed at the mRNA and protein levels through the activation of signal transducer and activator of transcription 5 (STAT5) in Ba/F3 cells expressing a JAK2V617F mutant and erythropoietin receptor (V617F/EpoR cells) and MPN patient-derived HEL cells. A treatment with the JAK1/2 inhibitor, ruxolitinib and STAT5 inhibitor, pimozide significantly inhibited DDX5 mRNA expression and enhanced the degradation of DDX5 in these cells, suggesting that the JAK2V617F mutant positively regulates DDX5 mRNA expression and DDX5 protein stability by activating STAT5. The knockdown of DDX5 specifically inhibited the activation of mechanistic target of rapamycin (mTOR) in V617F/EpoR cells and HEL cells and significantly suppressed the proliferation of these cells. Furthermore, the knockdown of DDX5 markedly suppressed tumorigenesis, splenomegaly, and liver hypertrophy caused by an inoculation of V617F/EpoR cells in nude mice. Collectively, these results revealed that JAK2V617F exhibits transforming activity by inducing the expression of DDX5 in a STAT5-dependent manner, indicating the potential of the JAK2V617F/STAT5/DDX5 axis as a therapeutic target in the treatment of MPN.

• Access to Document (DOI)

Identification of DDX5 as an indispensable activator of the glucocorticoid receptor in adipocyte differentiation.

Hokimoto S, Funakoshi-Tago M, Tago K.

FEBS J.  2022年09月

研究論文（学術雑誌）, 共著, 責任著者, 査読有り

• Access to Document (DOI)

Coffee ingredients, hydroquinone, pyrocatechol, and 4-ethylcatechol exhibit anti-inflammatory activity through inhibiting NF-κB and activating Nrf2

Funakoshi-Tago M., Matsutaka M., Hokimoto S., Kobata K., Tago K., Tamura H.

Journal of Functional Foods （Journal of Functional Foods）  90 2022年03月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  17564646

　概要を見る

Pyrocatechol is a coffee ingredient that exhibits anti-inflammatory activity. We herein identified hydroquinone and 4-ethylcatechol as novel coffee ingredients inhibiting LPS-induced inflammatory responses. Similar to pyrocatechol, hydroquinone and 4-ethylcatechol significantly inhibited LPS-induced nitric oxide (NO) production and CCL2 secretion by suppressing the mRNA expression of iNOS and CCL2 in murine macrophage-like RAW264.7 cells. These ingredients also inhibited the LPS-induced activation of NF-κB, a pivotal transcription factor in inflammation, by preventing its nuclear localization, and induced the expression of Nrf2, a transcription factor negatively regulating LPS-induced inflammation. By utilizing Keap1−/−MEFs reconstituted with Keap1 mutants in which major cysteine residues were substituted, we found that Cys151 in Keap1 was a critical sensor for pyrocatechol, hydroquinone, and 4-ethylcatechol to induce Nrf2 expression. However, the amounts of hydroquinone and 4-ethylcatechol in the coffee decoction were markedly lower than that of pyrocatechol, suggesting that pyrocatechol is the main coffee ingredient exhibiting anti-inflammatory activity.

• Access to Document (DOI)

A bis-pyridinium fullerene derivative induces apoptosis through the generation of ROS in BCR-ABL-positive leukemia cells

Sumi K., Tago K., Nakazawa Y., Takahashi K., Ohe T., Mashino T., Funakoshi-Tago M.

European Journal of Pharmacology （European Journal of Pharmacology）  916 2022年02月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  00142999

　概要を見る

A fusion protein, Breakpoint cluster region-Abelson (BCR-ABL) is responsible for the development of chronic myeloid leukemia (CML) and acute lymphocytic leukemia (ALL). Inhibitors against BCR-ABL are effective for the treatment of leukemia; however, a gatekeeper mutation (T315I) in BCR-ABL results in resistance to these inhibitors, which markedly impedes their efficacy. We herein demonstrated that a bis-pyridinium fullerene derivative (BPF) significantly induced apoptosis in human CML-derived K562 cells and ALL-derived SUP-B15 cells via the generation of reactive oxygen species (ROS). BPF reduced the expression of Bcr-Abl mRNA by inhibiting expression of c-Myc through ROS production. BPF also accelerated protein degradation of BCR-ABL through ROS production. Furthermore, BPF down-regulated the expression of not only BCR-ABL but also T315I-mutated BCR-ABL in ROS-dependent manner. As a result, BPF effectively induced apoptosis in transformed Ba/F3 cells expressing both BCR-ABL and T315I-mutated BCR-ABL. Collectively, these results indicate the potential of BPF as an effective leukemia drug that overcomes resistance to BCR-ABL inhibitors.

• Access to Document (DOI)

Novel Mechanism by a Bis-Pyridinium Fullerene Derivative to Induce Apoptosis by Enhancing the MEK-ERK Pathway in a Reactive Oxygen Species-Independent Manner in BCR-ABL-Positive Chronic Myeloid Leukemia-Derived K562 Cells

Sumi K., Tago K., Nakazawa Y., Takahashi K., Ohe T., Mashino T., Funakoshi-Tago M.

International Journal of Molecular Sciences 23 （ (2) ） 749 2022年01月

研究論文（学術雑誌）, 最終著者, 責任著者, 査読有り,  ISSN  16616596

　概要を見る

In the treatment of breakpoint cluster region-Abelson (BCR-ABL)-positive chronic myeloid leukemia (CML) using BCR-ABL inhibitors, the appearance of a gatekeeper mutation (T315I) in BCR-ABL is a serious issue. Therefore, the development of novel drugs that overcome acquired resistance to BCR-ABL inhibitors by CML cells is required. We previously demonstrated that a bis-pyridinium fullerene derivative (BPF) induced apoptosis in human chronic myeloid leukemia (CML)-derived K562 cells partially through the generation of reactive oxygen species (ROS). We herein show that BPF enhanced the activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase-extracellular signal-regulated kinase (MEK-ERK) pathway in a ROS-independent manner. BPF-induced apoptosis was attenuated by trametinib, suggesting the functional involvement of the MEK-ERK pathway in apoptosis in K562 cells. In addition, the constitutive activation of the MEK-ERK pathway by the enforced expression of the BRAFV600E mutant significantly increased the sensitivity of K562 cells to BPF. These results confirmed for the first time that BPF induces apoptosis in K562 cells through dual pathways—ROS production and the activation of the MEK-ERK pathway. Furthermore, BPF induced cell death in transformed Ba/F3 cells expressing not only BCR-ABL but also T315I mutant through the activation of the MEK-ERK pathway. These results indicate that BPF is as an effective CML drug that overcomes resistance to BCR-ABL inhibitors.

• Access to Document (DOI)

Identification of DDX5 as an indispensable activator of the glucocorticoid receptor in adipocyte differentiation

Hokimoto S., Funakoshi-Tago M., Tago K.

FEBS Journal （FEBS Journal）  2022年

ISSN  1742464X

　概要を見る

The expression of CCAAT/enhancer-binding protein (C/EBP) family members and peroxisome proliferator-activated receptor γ (PPAR γ) is essential for the differentiation of pre-adipocyte 3T3-L1 cells into mature adipocytes induced by a combined stimulation with dexamethasone, 3-isobutyl-1-methylxanthine and insulin (DMI). We herein demonstrated that the RNA helicase DDX5, the expression of which was induced by DMI, played an important role in the adipocyte differentiation of 3T3-L1 cells. The DMI-induced accumulation of lipid droplets and expression of adipocyte markers in 3T3-L1 cells were significantly inhibited by the knockdown of DDX5. The knockdown of DDX5 interfered with the expressional induction of C/EBPδ, which was the first to be induced in the transcription factor cascade, and inhibited the subsequent expression of the other transcription factors, C/EBPβ, PPARγ and C/EBPα. DDX5 interacted with the glucocorticoid receptor (GR), which induced the expression of C/EBPδ. The knockdown of DDX5 failed to induce the nuclear translocation of GR, suggesting the essential role of DDX5 in the early stage of adipocyte differentiation. Furthermore, the reconstitution of DDX5, but not the DDX5 mutant (K144N) lacking RNA helicase activity, restored DMI-induced GR activation and adipocyte differentiation in 3T3-L1 cells in which DDX5 was knocked down, confirming that the RNA helicase activity of DDX5 is essential for adipogenesis. Collectively, these results revealed for the first time that DDX5 is necessary for GR activation and plays an essential role in early adipocyte differentiation.

• Access to Document (DOI)

Capsaicin attenuates TGFβ2-induced epithelial-mesenchymal-transition in lens epithelial cells in vivo and in vitro

Sugiyama Y., Nakazawa Y., Sakagami T., Kawata S., Nagai N., Yamamoto N., Funakoshi-Tago M., Tamura H.

Experimental Eye Research （Experimental Eye Research）  213 2021年12月

研究論文（学術雑誌）, 査読有り,  ISSN  00144835

　概要を見る

Posterior capsule opacification (PCO), the most common complication of cataract surgery occurring in 20–50% of patients after 2–5 years of cataract surgery, is a major problem in the aging society. The epithelial-mesenchymal transition (EMT) of lens epithelial cells after cataract surgery has been proposed as a major cause of PCO. Capsaicin, widely used as a food additive and analgesic agent, is a major pungent ingredient in red pepper. Although the effect of capsaicin on EMT has been reported in cancer cells, the biological reaction of capsaicin was unique in each cell type, and there have been no reports describing its effects on EMT earlier. In this study, we demonstrated that treatment with capsaicin inhibited TGFβ2-induced EMT in vitro lens epithelial cells and ex vivo explant lens epithelial cells. Furthermore, eye drops of capsaicin inhibited the PCO model mice in vivo. Finally, we showed that capsaicin inhibited non-canonically induced Smad2/3 activation via suppression of EGFR activation and ERK phosphorylation. Our findings indicate that capsaicin and its derivatives are good candidate compounds for preventing PCO after cataract surgery.

• Access to Document (DOI)

K15 promoter-driven enforced expression of NKIRAS exhibits tumor suppressive activity against the development of DMBA/TPA-induced skin tumors

Tago K., Ohta S., Aoki-Ohmura C., Funakoshi-Tago M., Sashikawa M., Matsui T., Miyamoto Y., Wada T., Oshio T., Komine M., Matsugi J., Furukawa Y., Ohtsuki M., Yamauchi J., Yanagisawa K.

Scientific Reports （Scientific Reports）  11 （ 1 ）  2021年12月

研究論文（学術雑誌）, 共著, 査読有り

　概要を見る

NKIRAS1 and NKIRAS2 (also called as κB-Ras) were identified as members of the atypical RAS family that suppress the transcription factor NF-κB. However, their function in carcinogenesis is still controversial. To clarify how NKIRAS acts on cellular transformation, we generated transgenic mice in which NKIRAS2 was forcibly expressed using a cytokeratin 15 (K15) promoter, which is mainly activated in follicle bulge cells. The ectopic expression of NKIRAS2 was mainly detected in follicle bulges of transgenic mice with NKIRAS2 but not in wild type mice. K15 promoter-driven expression of NKIRAS2 failed to affect the development of epidermis, which was evaluated using the expression of K10, K14, K15 and filaggrin. However, K15 promoter-driven expression of NKIRAS2 effectively suppressed the development of skin tumors induced by treatment with 7,12-dimethylbenz(a)anthracene (DMBA)/12-O-tetradecanoylphorbol 13-acetate (TPA). This observation suggested that NKIRAS seemed to function as a tumor suppressor in follicle bulges. However, in the case of oncogenic HRAS-driven cellular transformation of murine fibroblasts, knockdown of NKIRAS2 expression drastically suppressed HRAS-mutant-provoked cellular transformation, suggesting that NKIRAS2 was required for the cellular transformation of murine fibroblasts. Furthermore, moderate enforced expression of NKIRAS2 augmented oncogenic HRAS-provoked cellular transformation, whereas an excess NKIRAS2 expression converted its functional role into a tumor suppressive phenotype, suggesting that NKIRAS seemed to exhibit a biphasic bell-shaped enhancing effect on HRAS-mutant-provoked oncogenic activity. Taken together, the functional role of NKIRAS in carcinogenesis is most likely determined by not only cellular context but also its expression level.

• Access to Document (DOI)

The indole-hydantoin derivative exhibits anti-inflammatory activity by preventing the transactivation of NF-κB through the inhibition of NF-κB p65 phosphorylation at Ser276

Lin X., Tago K., Okazaki N., So T., Takahashi K., Mashino T., Tamura H., Funakoshi-Tago M.

International Immunopharmacology （International Immunopharmacology）  100 2021年11月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  15675769

　概要を見る

Indole- and hydantoin-based derivatives both exhibit anti-inflammatory activity, suggesting that the structures of indole and hydantoin are functional for this activity. In the present study, we synthesized two types of indole-hydantoin derivatives, IH-1 (5-(1H-indole-3-ylmethylene) imidazolidine-2,4-dione) and IH-2 (5-(1H-indole-3-ylmethyl) imidazolidine-2,4-dione) and examined their effects on LPS-induced inflammatory responses in murine macrophage-like RAW264.7 cells. LPS-induced inflammatory responses were not affected by indole, hydantoin, or IH-2. In contrast, IH-1 significantly inhibited the LPS-induced production of nitric oxide (NO) and secretion of CCL2 and CXCL1 by suppressing the mRNA expression of inducible NO synthase (iNOS), CCL2, and CXCL1. IH-1 markedly inhibited the LPS-induced activation of NF-κB without affecting the degradation of IκBα or nuclear translocation of NF-κB. IH-1 markedly attenuated the transcriptional activity of NF-κB by suppressing the LPS-induced phosphorylation of the NF-κB p65 subunit at Ser276. Furthermore, IH-1 prevented the LPS-induced interaction of NF-κB p65 subunit with a transcriptional coactivator, cAMP response element-binding protein (CBP). Collectively, these results revealed the potential of the novel indole-hydantoin derivative, IH-1 as an anti-inflammatory drug.

• Access to Document (DOI)

Oral consumption of α-glucosyl-hesperidin could prevent lens hardening, which causes presbyopia

Nakazawa Y., Aoki M., Doki Y., Morishita N., Endo S., Nagai N., Funakoshi-Tago M., Tamura H.

Biochemistry and Biophysics Reports （Biochemistry and Biophysics Reports）  25 2021年03月

研究論文（学術雑誌）, 共著, 査読有り

　概要を見る

© 2020 The Authors Presbyopia is one of the most well-known diseases of the eye, predominantly affecting the adult population after 50 years’. Due to hardening of the lens and failure of accommodative change, patients lose the ability to focus on near objects. This eye symptom is reported to be an early symptom of age-related nuclear cataract, and we have previously reported that hesperetin treatment could delay the onset of nuclear cataractogenesis induced by sodium selenite. In this study, we examined whether oral intake of α-glucosyl-hesperidin (G-Hsd), which has greater water solubility than hesperetin, could delay the onset of presbyopia. G-Hsd treatment protected lens elasticity, upregulated the mRNA expression of anti-oxidative enzymes like glutathione reductase and thioredoxin reductase 1 in the plasma and lens, and prevented premature cataract symptoms in selenite-induced cataract rat lens. Thus, the anti-presbyopic effects of G-Hsd were attributed, at least in part, to its antioxidant effects. G-Hsd represents the first oral treatment agent with anti-presbyopia and/or anti-cataract properties.

• Access to Document (DOI)

Effect of alpha‐glucosyl‐hesperidin consumption on lens sclerosis and presbyopia

Nakazawa Y., Doki Y., Sugiyama Y., Kobayashi R., Nagai N., Morisita N., Endo S., Funakoshi‐tago M., Tamura H.

Cells （Cells）  10 （ 2 ） 1 - 11 2021年02月

研究論文（学術雑誌）, 共著, 査読有り

　概要を見る

Presbyopia is characterized by a decline in the ability to accommodate the lens. The most commonly accepted theory for the onset of presbyopia is an age‐related increase in the stiffness of the lens. However, the cause of lens sclerosis remains unclear. With age, water microcirculation in the lens could change because of an increase in intracellular pressure. In the lens, the intracellular pressure is controlled by the Transient Receptor Potential Vanilloid (TRPV) 1 and TRPV4 feedback pathways. In this study, we tried to elucidate that administration of α‐glucosyl‐hesperidin (G‐Hsd), previously reported to prevent nuclear cataract formation, affects lens elasticity and the distribution of TRPV channels and Aquaporin (AQP) channels to meet the requirement of intracellular pressure. As a result, the mouse control lens was significantly toughened compared to both the 1% and 2% G‐Hsd mouse lens treatments. The anti‐oxidant levels in the lens and plasma decreased with age; however, this decrease could be nullified with either 1% or 2% G‐Hsd treatment in a concentration-and exposure time‐dependent manner. Moreover, G‐Hsd treatment affected the TRPV4 distribu-tion, but not TRPV1, AQP0, and AQP5, in the peripheral area and could maintain intracellular pres-sure. These findings suggest that G‐Hsd has great potential as a compound to prevent presbyopia and/or cataract formation.

• Access to Document (DOI)

EBP2, a novel NPM-ALK-interacting protein in the nucleolus, contributes to the proliferation of ALCL cells by regulating tumor suppressor p53

Uchihara Y., Tago K., Tamura H., Funakoshi-Tago M.

Molecular Oncology （Molecular Oncology）  15 （ 1 ） 167 - 197 2021年01月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  15747891

　概要を見る

The oncogenic fusion protein nucleophosmin-anaplastic lymphoma kinase (NPM-ALK), found in anaplastic large-cell lymphoma (ALCL), localizes to the cytosol, nucleoplasm, and nucleolus. However, the relationship between its localization and transforming activity remains unclear. We herein demonstrated that NPM-ALK localized to the nucleolus by binding to nucleophosmin 1 (NPM1), a nucleolar protein that exhibits shuttling activity between the nucleolus and cytoplasm, in a manner that was dependent on its kinase activity. In the nucleolus, NPM-ALK interacted with Epstein–Barr virus nuclear antigen 1-binding protein 2 (EBP2), which is involved in rRNA biosynthesis. Moreover, enforced expression of NPM-ALK induced tyrosine phosphorylation of EBP2. Knockdown of EBP2 promoted the activation of the tumor suppressor p53, leading to G0/G1-phase cell cycle arrest in Ba/F3 cells transformed by NPM-ALK and ALCL patient-derived Ki-JK cells, but not ALCL patient-derived SUDH-L1 cells harboring p53 gene mutation. In Ba/F3 cells transformed by NPM-ALK and Ki-JK cells, p53 activation induced by knockdown of EBP2 was significantly inhibited by Akt inhibitor GDC-0068, mTORC1 inhibitor rapamycin, and knockdown of Raptor, an essential component of mTORC1. These results suggest that the knockdown of EBP2 triggered p53 activation through the Akt-mTORC1 pathway in NPM-ALK-positive cells. Collectively, the present results revealed the critical repressive mechanism of p53 activity by EBP2 and provide a novel therapeutic strategy for the treatment of ALCL.

• Access to Document (DOI)

The role of MerTK in promoting cell migration is enhanced by the oncogenic Ras/IL-33 signaling axis

Ohta S., Tago K., Kuchimaru T., Funakoshi-Tago M., Horie H., Aoki-Ohmura C., Matsugi J., Yanagisawa K.

FEBS Journal （FEBS Journal）  289 （ 7 ） 1950 - 1967 2021年

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  1742464X

　概要を見る

Ras genes are frequently mutated in many cancer types; however, there are currently no conclusively effective anticancer drugs against Ras-induced cancer. Therefore, the downstream effectors of Ras signaling need to be identified for the development of promising novel therapeutic approaches. We previously reported that oncogenic Ras induced the expression of NF-HEV/IL-33, a member of the interleukin-1 family, and showed that intracellular IL-33 was required for oncogenic Ras-induced cellular transformation. In the present study, we demonstrated that the c-Mer proto-oncogene tyrosine kinase (MerTK), a receptor tyrosine kinase, played essential roles in oncogenic Ras/IL-33 signaling. The expression of MerTK was enhanced in transformed NIH-3T3 cells by the expression of oncogenic Ras, H-Ras (G12V), in an IL-33-dependent manner. In human colorectal cancer tissues, MerTK expression also correlated with IL-33 expression. The knockdown of IL-33 or MerTK effectively attenuated the migration of NIH-3T3 cells transformed by H-Ras (G12V) and A549, LoVo, and HCT116 cells harboring an oncogenic K-Ras mutation. Furthermore, the suppression of Ras-induced cell migration by the knockdown of IL-33 was rescued by the enforced expression of MerTK. The present results also revealed that MerTK was effectively phosphorylated in NIH-3T3 cells transformed by Ras (G12V). Ras signaling was essential for the tyrosine phosphorylation of MerTK, and the kinase activity of MerTK was indispensable for accelerating cell migration. Collectively, the present results reveal a novel role for MerTK in cancer malignancy, which may be utilized to develop novel therapeutic strategies that target Ras-transformed cells.

• Access to Document (DOI)

Pyrocatechol, a component of coffee, suppresses LPS-induced inflammatory responses by inhibiting NF-κB and activating Nrf2

Funakoshi-Tago M., Nonaka Y., Tago K., Takeda M., Ishihara Y., Sakai A., Matsutaka M., Kobata K., Tamura H.

Scientific Reports （Scientific Reports）  10 （ 1 ）  2020年12月

研究論文（学術雑誌）, 共著, 査読有り

　概要を見る

© 2020, The Author(s). Coffee is a complex mixture of many bioactive compounds possessing anti-inflammatory properties. However, the mechanisms by which coffee exerts anti-inflammatory effects remains unclear and the active ingredients have not yet been identified. In this study, we found that coffee extract at more than 2.5%(v/v) significantly inhibited LPS-induced inflammatory responses in RAW264.7 cells and that anti-inflammatory activity of coffee required the roasting process. Interestingly, we identified pyrocatechol, a degradation product derived from chlorogenic acid during roasting, as the active ingredient exhibiting anti-inflammatory activity in coffee. HPLC analysis showed that 124 μM pyrocatechol was included in 100% (v/v) roasted coffee. A treatment with 5%(v/v) coffee extract and more than 2.5 μM pyrocatechol inhibited the LPS-induced activation of NF-κB and also significantly activated Nrf2, which acts as a negative regulator in LPS-induced inflammation. Furthermore, intake of 60% (v/v) coffee extract and 74.4 μM pyrocatechol, which is the concentration equal to contained in 60% (v/v) coffee, markedly inhibited the LPS-induced inflammatory responses in mice. Collectively, these results demonstrated that pyrocatechol, which was formed by the roasting of coffee green beans, is one of the ingredients contributing to the anti-inflammatory activity of coffee.

• Access to Document (DOI)

Coffee decoction enhances tamoxifen proapoptotic activity on MCF-7 cells

Funakoshi-Tago M., Tago K., Li C., Hokimoto S., Tamura H.

Scientific Reports （Scientific Reports）  10 （ 1 ） 19588 2020年11月

研究論文（学術雑誌）, 共著, 査読有り

　概要を見る

The consumption of coffee has been suggested to effectively enhance the therapeutic effects of tamoxifen against breast cancer; however, the underlying molecular mechanisms remain unclear. We herein attempted to clarify how coffee decoction exerts anti-cancer effects in cooperation with tamoxifen using the estrogen receptor α (ERα)-positive breast cancer cell line, MCF-7. The results obtained showed that coffee decoction down-regulated the expression of ERα, which was attributed to caffeine inhibiting its transcription. Coffee decoction cooperated with tamoxifen to induce cell-cycle arrest and apoptotic cell death, which may have been mediated by decreases in cyclin D1 expression and the activation of p53 tumor suppressor. The inclusion of caffeine in coffee decoction was essential, but not sufficient, to induce cell-cycle arrest and apoptotic cell death, suggesting the requirement of unknown compound(s) in coffee decoction to decrease cyclin D1 expression and activate apoptotic signaling cascades including p53. The activation of p53 through the cooperative effects of these unidentified component(s), caffeine, and tamoxifen appeared to be due to the suppression of the ERK and Akt pathways. Although the mechanisms by which the suppression of these pathways induces p53-mediated apoptotic cell death remain unclear, the combination of decaffeinated coffee, caffeine, and tamoxifen also caused cell-cycle arrest and apoptotic cell death, suggesting that unknown compound(s) present in decaffeinated coffee cooperate with caffeine and tamoxifen.

• Access to Document (DOI)

Kampo medicines, Rokumigan, Hachimijiogan, and Goshajinkigan, significantly inhibit glucagon-induced CREB activation

Funakoshi-Tago M., Yu S., Kushida A., Takeuchi K., Tamura H.

Heliyon （Heliyon）  6 （ 3 ）  2020年03月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  24058440

　概要を見る

© 2020 The Author(s) The pathophysiology of type 2 diabetes mellitus (T2DM) is characterized by not only insulin resistance, but also the abnormal regulation of glucagon secretion, suggesting that antagonizing the glucagon-induced signaling pathway has therapeutic potential in the treatment of T2DM. Although various Kampo medicines (traditional herbal medicines) are often utilized to ameliorate the symptoms of T2DM, their effects on glucagon signaling have not yet been clarified. In the present study, we examined the effects of nine types of representative Kampo formulations prescribed for T2DM on glucagon-induced CREB activation in HEK293T cells stably expressing glucagon receptor (Gcgr) and a hepatic cell line HepG2. Among these Kampo medicines, Rokumigan, Hachimijiogan, and Goshajinkigan significantly suppressed the glucagon-induced transactivation of the cAMP-responsive element (CRE)-binding protein (CREB) by inhibiting its interaction with CREB-binding protein (CBP), which led to a reduction in the expression of phosphoenolpyruvate carboxykinase (PEPCK) mRNA. Furthermore, among the crude drugs commonly contained in these three Kampo medicines, Rehmannia Root (Jio), Moutan Bark (Botampi), and Cornus Fruit (Shanzhuyu) exerted inhibitory effects on glucagon-induced CREB activation. Collectively, the present results provide a novel mechanism, the inhibition of glucagon signaling, by which Rokumigan, Hachimijiogan, and Goshajinkigan improve the symptoms of T2DM.

• Access to Document (DOI)

Oral intake of α-glucosyl-hesperidin ameliorates selenite-induced cataract formation

Nakazawa Y., Aoki M., Ishiwa S., Morishita N., Endo S., Nagai N., Yamamoto N., Funakoshi-Tago M., Tamura H.

Molecular Medicine Reports （Molecular Medicine Reports）  21 （ 3 ） 1258 - 1266 2020年

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  17912997

　概要を見る

© 2020 Spandidos Publications. All rights reserved. Hesperetin is a natural flavonoid with robust antioxidant properties. Our previous study reported that hesperetin can prevent cataract formation. However, an important consideration regarding hesperetin consumption is the limited bioavailability due to its poor solubility. The present study investigated the anti-cataract effects of α-glucosyl hesperidin in vivo and in vitro using a selenite-induced cataract model. SD rats (age, 13 days) were orally administered PBS (0.2 ml) or α-glucosyl hesperidin (200 mg/kg) on days 0, 1 and 2. Sodium selenite was subcutaneously administered to the rats 4 h after the first oral administration on day 0. Antioxidant levels in the lens and blood were measured on day 6. In vitro, human lens epithelial cells were treated with sodium selenite (10 μM) and/or hesperetin (50 or 100 mM) for 24 h and analyzed for apoptosis markers using sub-G1 population and Annexin V-FITC/propidium iodide staining and DNA ladder formation. α-glucosyl hesperidin treatment significantly reduced the severity of selenite-induced cataract. The level of antioxidants was significantly reduced in the selenite-treated rats compared with in the controls; however, they were normalized with α-glucosyl hesperidin treatment. In vitro, hesperetin could significantly reduce the number of cells undergoing apoptosis induced by sodium selenite in human lens epithelial cell lines. Overall, oral consumption of α-glucosyl hesperidin could delay the onset of selenite-induced cataract, at least in part by modulating the selenite-induced cell death in lens epithelial cells.

• Access to Document (DOI)

N-Acetyl cysteine prevents activities of STAT3 inhibitors, Stattic and BP-1-102 independently of its antioxidant properties

Uchihara Y., Ohe T., Mashino T., Kidokoro T., Tago K., Tamura H., Funakoshi-Tago M.

Pharmacological Reports （Pharmacological Reports）  71 （ 6 ） 1067 - 1078 2019年12月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  17341140

　概要を見る

© 2019 Institute of Pharmacology, Polish Academy of Sciences Background: Inhibitors for signal transducer and activator of transcription 3 (STAT3), Stattic, BP-1-102, and LLL12 significantly induce apoptosis in transformed Ba/F3 cells expressing an oncogenic fusion protein, nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) that induces the activation of STAT3. We found that the antioxidant reagent, N-acetyl cysteine (NAC) prevented the abilities of Stattic and BP-1-102, but not LLL12 to induce apoptosis in transformed cells expressing NPM-ALK, providing a novel problem in use of STAT3 inhibitors. We herein investigated the mechanisms how NAC prevented the effects of Sttatic and BP-1-102. Methods: Ba/F3 cells expressing NPM-ALK and SUDHL-1 cells were treated with antioxidants such as NAC, Trolox or edaravone in combination with STAT3 inhibitors. Phosphorylation of STAT3, cell proliferation rate, cell viability, cell cycle, internucleosomal DNA fragmentation and the intracellular accumulation of reactive oxygen species (ROS) was investigated. The binding of STAT3 inhibitors and NAC was analyzed by LC–MS. Results: NAC but not Trolox and edaravone diminished the abilities of Stattic and BP-1-102 to induce apoptosis in cells expressing NPM-ALK. The ROS levels in cells expressing NPM-ALK were not markedly affected by the treatments with Stattic and BP-1-102 in combination with NAC, suggesting that NAC inhibited the activity of Stattic and BP-1-102 independent of its antioxidant activity. LC–MS analysis revealed that NAC directly bound to Stattic and BP-1-102. Furthermore, these NAC adducts exhibited no cytotoxicity, and failed to affect the activity of STAT3. Conclusions: NAC antagonizes the activities of Stattic and BP-1-102, which inhibit STAT3 activation by interacting with cysteine residues in STAT3.

• Access to Document (DOI)

Methotrexate significantly induces apoptosis by inhibiting STAT3 activation in NPM-ALK-positive ALCL cells

Uchihara Y., Komori R., Tago K., Tamura H., Funakoshi-Tago M.

Biochemical Pharmacology （Biochemical Pharmacology）  170 2019年12月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  00062952

　概要を見る

© 2019 Elsevier Inc. Anaplastic large cell lymphoma (ALCL) is associated with a characteristic chromosomal translocation that generates the oncogenic fusion protein, nucleophosmin-anaplastic lymphoma kinase (NPM-ALK). Methotrexate is a commonly used chemotherapeutic drug in the treatment of multiple cancers due to its inhibition of dihydrofolate reductase (DHFR), which suppresses the synthesis of DNA. In the present study, we found that low-dose methotrexate significantly induced apoptosis in transformed Ba/F3 cells expressing NPM-ALK by inhibiting the activation of signal transducer and activator of transcription factor 3 (STAT3), a critical downstream molecule of NPM-ALK. Although methotrexate prevented the phosphorylation of STAT3, it did not affect the activity of NPM-ALK. A co-treatment with folinic acid prevented the methotrexate-induced inhibition of STAT3 activation and induction of apoptosis, suggesting that methotrexate exerts its cytotoxic effects by depleting tetrahydrofolate (THF) in transformed cells by NPM-ALK. Furthermore, methotrexate induced the down-regulation of the anti-apoptotic protein, MCL-1, DNA damage, and the activation of a p53 tumor suppressor, leading to apoptosis through the inhibition of STAT3. Methotrexate significantly induced apoptosis in ALK inhibitor-resistant cells expressing the NPM-ALK mutant harboring the point mutation, G262R, and in ALCL patient-derived NPM-ALK-positive Ki-JK cells. Collectively, these results demonstrate the potential therapeutic application of methotrexate, which inhibits the activation of STAT3, to NPM-ALK-positive ALCL.

• Access to Document (DOI)

Oncogenic Ras mutant causes the hyperactivation of NF-κB via acceleration of its transcriptional activation

Tago K., Funakoshi-Tago M., Ohta S., Kawata H., Saitoh H., Horie H., Aoki-Ohmura C., Yamauchi J., Tanaka A., Matsugi J., Yanagisawa K.

Molecular Oncology （Molecular Oncology）  13 （ 11 ） 2493 - 2510 2019年11月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  15747891

　概要を見る

© 2019 The Authors. Published by FEBS Press and John Wiley & Sons Ltd. It is well established that nuclear factor κB (NF-κB) acts as one of the most important transcription factors for tumor initiation and progression, as it both protects cells from apoptotic/necrotic signals and accelerates angiogenesis and tumor metastasis, which is mediated via the expression of target genes. However, it has not yet been clarified how oncogenic signals accelerate the activation of NF-κB. In the current study, we utilized untransformed NIH-3T3 cells stably harboring a κB-driven luciferase gene to show that an oncogenic mutant of Ras GTPase augmented TNFα-induced NF-κB activation. Notably, enforced expression of cyclin-dependent kinase inhibitors, such as p27Kip1 and p21Cip1, effectively canceled the accelerated activation of NF-κB, suggesting that oncogenic Ras-induced cell cycle progression is essential for the hyperactivation of NF-κB. Furthermore, we found that Ras (G12V) augmented the transcriptional activation of NF-κB, and this activation required the p38 MAP kinase. We observed that a downstream kinase of p38 MAP kinase, MSK1, was activated by Ras (G12V) and catalyzed the phosphorylation of p65/RelA at Ser-276, which is critical for its transcriptional activation. Significantly, phosphorylation of the p65/RelA subunit at Ser-276 was elevated in patient samples of colorectal cancer harboring oncogenic mutations of the K-Ras gene, and the expression levels of NF-κB target genes were drastically enhanced in several cancer tissues. These observations strongly suggest that oncogenic signal-induced acceleration of NF-κB activation is caused by activation of the p38 MAP kinase–MSK1 signaling axis and by cell cycle progression in cancer cells.

• Access to Document (DOI)

Tyrosine-phosphorylated SOCS3 negatively regulates cellular transformation mediated by the myeloproliferative neoplasm-associated JAK2 V617F mutant

Funakoshi-Tago M., Tsuruya R., Ueda F., Ishihara A., Kasahara T., Tamura H., Tago K.

Cytokine （Cytokine）  123 2019年11月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  10434666

　概要を見る

© 2019 Elsevier Ltd In the majority of myeloproliferative neoplasms (MPNs) patients, a point mutation, V617F has been found in Janus kinase 2 (JAK2) gene, and this JAK2 mutant provoked aberrant signaling pathway. In the current study, we found that suppressor of cytokine signaling proteins 3 (SOCS3) possessed the tumor suppressive activity against the JAK2 V617F mutant-provoked cellular transformation. The knockdown of SOCS3 increased the expression level of the JAK2 V617F mutant, which enhanced the activation of signaling mediators, including signal transducer and activator of transcription 3 and 5 (STAT3, STAT5) and extracellular signal-regulated kinase (ERK), and also increased of the proliferation rate and tumorigenesis activity of Ba/F3 cells expressing the JAK2 V617F mutant and erythropoietin receptor (EpoR). In contrast, the enforced expression of SOCS3 significantly inhibited the JAK2 V617F mutant-induced activation of downstream signaling molecules, cell proliferation, and tumorigenesis by down-regulating the expression level of the JAK2 V617F mutant. SOCS3 interacted with the JAK2V617F mutant through its SH2 domain and was phosphorylated at Tyr-204 and Tyr-221 in its SOCS box by the JAK2V617F mutant. SOCS3 mutants carrying a mutation in the SH2 domain (R71E) and a substitution at Tyr-221 (Y221F) failed to exert inhibitory effects on JAK2V617F mutant-induced cellular transformation and tumorigenesis. Collectively, these results imply that SOCS3 plays a negative role in the JAK2 V617F mutant-induced oncogenic signaling pathway through its SH2 domain and the phosphorylation of Tyr-221 in its SOCS box.

• Access to Document (DOI)

The mechanisms of taxodione-induced apoptosis in BCR-ABL-positive leukemia cells

Uchihara Y., Tago K., Funakoshi-Tago M.

Folia Pharmacologica Japonica （Folia Pharmacologica Japonica）  153 （ 4 ） 147 - 154 2019年

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  00155691

　概要を見る

© 2019, Japanese Pharmacological Society. All rights reserved. Chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL) are caused by a fusion protein, BCR-ABL, which induces cellular transformation by activating the signaling molecules, STAT5 and Akt. The specific BCR-ABL inhibitors including imatinib, nilotinib, and dasatinib, are clinically utilized in the treatment with CML and ALL patients. Although these BCR-ABL inhibitors are initially successful in the treatment of leukemia, many patients develop drug resistance due to the appearance of the gatekeeper mutation of BCR-ABL, T315I. Recently, we found that taxodione, a quinone methide diterpene isolated from a conifer, Taxodium distichum, significantly induced apoptosis in human myelogenous leukemia-derived K562 cells, which is positive for the bcr-abl gene. Taxodione reduced the activities of mitochondrial respiratory chain complex III, leading to the production of reactive oxygen species (ROS). An antioxidant agent, N-acetylcysteine (NAC), canceled taxodione-induced ROS production and apoptotic cell death, suggesting that taxodione induced apoptosis through ROS accumulation. Furthermore, in K562 cells treated with taxodione, BCR-ABL, STAT5 and Akt were sequestered in mitochondrial fraction, and their localization changes decrease their abilities to stimulate cell proliferation. Strikingly, NAC canceled these taxodione-caused inhibition of BCR-ABL, STAT5 and Akt. In addition, taxodione significantly induced apoptosis in transformed Ba/F3 cells by not only BCR-ABL but also T315I-mutated BCR-ABL through the generation of ROS, suggesting that taxodione has potential as anti-tumor drug with high efficacy to overcome BCR-ABL T315I mutation-mediated resistance in leukemia cells. It’s also expected that these knowledge becomes an important clue in the development of anti-cancer drugs against the broad range of tumors.

• Access to Document (DOI)

Coffee brew intake can prevent the reduction of lens glutathione and ascorbic acid levels in hfd-fed animals

多胡 めぐみ

Experimental and Therapeutic Medicine （Experimental and Therapeutic Medicine）  17 （ 2 ） 1420 - 1425 2018年12月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  17920981

　概要を見る

© 2018, Spandidos Publications. All rights reserved. The lens has high concentrations of glutathione (GSH) and ascorbic acid (AsA) to maintain redox activity and prevent cataract formation, which is the leading cause of visual impairment worldwide. Metabolic syndrome is reported to be linked with a higher risk of age-associated cataract. As it was demonstrated previously that coffee consumption improved high-fat diet (HFD) -induced metabolic symptoms, it was hypothesized that coffee intake could delay the onset of obesity related-cataract; however, the effect of coffee consumption on this type of cataract remains unknown. Four-week-old male C57BL/6JJms SLC mice were divided into two groups and were provided ad libitum access to either a control diet (control groups) or a HFD (HFD groups). The control groups and HFD groups were further divided into three or four subgroups for each experiment. Coffee intake markedly reduced the increase in body weight in a roasting-time and concentration-dependent manner. Coffee consumption also prevented the HFD-induced decrease in the concentration of GSH and AsA, and treatment with pyrocatechol or caffeine also restored the reduction of antioxidant compounds. Plasma cholesterol and triglycerides were significantly higher in HFD groups; however, coffee brew or coffee constituent treatment in the HFD-fed mice group prevented elevation of these levels. Caffeine is a major coffee component and pyrocatechol is generated thought the roasting process. These results revealed that caffeine and pyrocatechol in coffee brew may be the key constituents responsible for preventing the reduction of lens GSH and AsA in HFD-fed animals.

• Access to Document (DOI)

Chemoenzymatic synthesis of hydroxytyrosol monoesters and their suppression effect on nitric oxide production stimulated by lipopolysaccharides

多胡 めぐみ

Bioscience, Biotechnology and Biochemistry （Bioscience, Biotechnology and Biochemistry）  83 （ 2 ） 185 - 191 2018年10月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  09168451

　概要を見る

© 2018 Japan Society for Bioscience, Biotechnology, and Agrochemistry. Fatty acid monoesters of hydroxytyrosol [2-(3,4-dihydroxyphenyl)ethanol] were synthesized in two steps from tyrosol (4-hydroxyphenylethanol) by successive Candida antarctica lipase B-catalyzed chemoselective acylation on the primary aliphatic hydroxy group over phenolic hydroxy group in tyrosol, and 2-iodoxybenzoic acid (IBX)-mediated hydroxylation adjacent to the remaining free phenolic hydroxy group. Examination of their suppression effects on nitric oxide production stimulated by lipopolysaccharides in RAW264.7 cells showed that hydroxytyrosol butyrate exhibited the highest inhibition (IC 50 7.0 μM) among the tested compounds.

• Access to Document (DOI)

Roasted Coffee Reduces β-Amyloid Production by Increasing Proteasomal β-Secretase Degradation in Human Neuroblastoma SH-SY5Y Cells

多胡 めぐみ

Molecular Nutrition and Food Research （Molecular Nutrition and Food Research）  62 （ 21 ） e1800238 2018年09月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  16134125

　概要を見る

© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim Scope: Epidemiological studies have shown that coffee consumption may be associated with a lower risk of developing several neurological disorders, including Alzheimer's disease (AD). Caffeine is a prominent candidate component underlying the preventive effects of coffee; however, the contribution of other constituents is unclear. To clarify this issue, the effect of roasting coffee beans on β-secretase (BACE1) expression in human neuroblastoma SH-SY5Y cells is investigated. Methods and results: Coffee (2%) reduces Aβ accumulation in culture medium to 80% of control levels after 24 h. Accordingly, BACE1 expression is decreased to 70% of control levels at 12 h. Experiments using cycloheximide and MG132, a proteasome inhibitor, reveal that coffee enhanced BACE1 degradation through activation of proteasomal activity. Furthermore, coffee activates cAMP-dependent protein kinase, and consequently, phosphorylation of a serine residue of proteasome 26S subunit, non-ATPase 11 (PSMD11). Pyrocatechol, a strong antioxidant known as catechol or 1,2-dihydroxybenzene, produced from chlorogenic acid during roasting, also reduces BACE1 expression by activation of proteasomal activity. Furthermore, pyrocatechol reduces Aβ production in SH-SY5Y cells. Conclusion: The data suggest that the roasting process may be crucial for the protective effects of coffee consumption in AD.

• Access to Document (DOI)

Hydroxytyrosol butyrate inhibits 6-OHDA-induced apoptosis through activation of the Nrf2/HO-1 axis in SH-SY5Y cells

多胡 めぐみ

European Journal of Pharmacology （European Journal of Pharmacology）  834   246 - 256 2018年07月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  00142999

　概要を見る

© 2018 Elsevier B.V. Hydroxytyrosol (HT) is a polyphenol contained in olives and exhibits antioxidant activity. We herein investigated the effects of HT and its derivatives, hydroxytyrosol acetate (HT-A) and hydroxytyrosol butyrate (HT-B), on the protection of neuronal cells against apoptosis induced by the Parkinson's disease-related neurotoxin 6-hydroxydopamine (6-OHDA). The pretreatment of SH-SY5Y cells with HT-B, but not HT or HT-A significantly reduced the 6-OHDA-induced generation of reactive oxygen species, activation of caspase-3, and subsequent cell death. HT-B also induced the protein expression of the transcription factor, NF-E2-related factor-2 (Nrf2) and its transcriptional activation, resulting in the up-regulated expression of heme oxygenase-1 (HO-1), which conferred neuroprotection against 6-OHDA-induced oxidative damage. Furthermore, three cysteine residues, Cys151, Cys273, and Cys288 in Kelch-like ECH-associated protein 1 (Keap1) were necessary for the HT-B-induced activation of Nrf2. Collectively, the present results demonstrated that HT-B, harboring higher fat solubility than HT and HT-A, effectively elicited adaptive responses to oxidative stress by activating the Nrf2/HO-1 axis in neuronal cells.

• Access to Document (DOI)

A 5-hydroxyoxindole derivative attenuates LPS-induced inflammatory responses by activating the p38-Nrf2 signaling axis

多胡 めぐみ

Biochemical Pharmacology （Biochemical Pharmacology）  155   182 - 197 2018年06月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  00062952

　概要を見る

© 2018 Elsevier Inc. 5-Hydroxyoxindole is a urinary metabolite of indole that exhibits antioxidant activity. In the present study, we found that a 5-hydroxyoxindole derivative (5-HI) significantly inhibited LPS-induced inflammatory effects in the murine macrophage cell line, RAW264.7. 5-HI induced the expression of the transcription factor, Nrf2, which is typically ubiquitinated by Keap1, an adaptor component of the ubiquitin E3 ligase complex, resulting in its proteasomal degradation. By utilizing Keap1−/− MEFs reconstituted with Keap1 mutants harboring substitutions in their major cysteine residues, we clarified the importance of Cys151 in Keap1 as a sensor for 5-HI in the induction of Nrf2 expression. Furthermore, 5-HI induced the activation of the MKK3/6-p38 pathway, which is required for the transcriptional activation of Nrf2. The knockdown of Nrf2 enhanced the LPS-induced expression of inflammatory mediators, including iNOS, NO, and CCL2, and effectively repressed the inhibitory effects of 5-HI on their expression. Although 5-HI and antioxidant N-acetyl cysteine (NAC) both reduced LPS-induced ROS generation, the treatment with NAC did not affect the LPS-induced expression of inflammatory mediators, suggesting that the anti-inflammatory activity of 5-HI mediated by Nrf2 is independent of redox control. Furthermore, when injected into mice with 5-HI, the expression of Nrf2 was significantly increased, and the LPS-induced mRNA expression of CXCL1, CCL2, TNFα and IL-6 were remarkably inhibited in the kidneys, liver, and lungs, and the production of these cytokines in serum was effectively reduced. Collectively, these results suggest that 5-HI has potential in the treatment of inflammatory diseases through the activation of Nrf2.

• Access to Document (DOI)

Effect of hesperetin derivatives on the development of selenite-induced cataracts in rats

多胡 めぐみ

Molecular Medicine Reports （Molecular Medicine Reports）  18 （ 1 ） 1043 - 1050 2018年05月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  17912997

　概要を見る

Copyright © Spandidos Publications 2017. All rights reserved. Cataracts are a major cause of blindness worldwide. As anti-cataract pharmaceutical therapies require long-term treatment, identifying anti-cataract compounds that are ubiquitous in the human diet, have no adverse effects and are affordable, is of paramount importance. The present study focused on hesperetin and its derived compounds, hesperetin stearic acid ester (Hes-S) and hesperetin oleic acid ester (Hes-O), in order to investigate their therapeutic potential to treat cataracts in a selenite animal model. Thirteen-day-old Sprague Dawley rats were divided into 12 groups. Animals in groups 1 and 7 were subcutaneously injected with vehicle, those in groups 2 and 8 were administered hesperetin, those in groups 3 and 9 received stearic acid, those in groups 4 and 10 were injected with oleic acid, those in groups 5 and 11 were administered Hes-S, and those in groups 6 and 12 received Hes-O (10 nmol/kg body weight on days 0, 1 and 2). Animals in groups 7 to 12 were treated with sodium selenite (20 µmol/kg body weight given 4 h following the test compound treatment on day 0) to induce cataract. On day 6, rats had less severe central opacities and lower stage cataracts than rats in the selenite treatment-only control groups. The levels of glutathione (GSH) and ascorbic acid (AsA) in lenses with selenite-induced cataracts declined to one-third of that of controls, and the reduction in GSH and AsA levels was rescued following hesperetin, Hes-S or Hes-O treatment, with concentrations remaining to 70-80% of that of controls. However, there were no changes in the plasma levels of GSH and AsA following treatments. Administration of either hesperetin or hesperetin-derived compounds prevented the reduction of chaperone activity in the lens, and rats treated with Hes-S or Hes-O treatment had significantly greater chaperone activity than hesperetin-treated rats. Collectively, these results suggested that hesperetin and hesperetin-derived compounds may be novel drug compounds that have the potential to prevent or delay the onset of cataracts.

• Access to Document (DOI)

Taxodione induces apoptosis in BCR-ABL-positive cells through ROS generation

多胡 めぐみ

Biochemical Pharmacology （Biochemical Pharmacology）  154   357 - 372 2018年05月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  00062952

　概要を見る

© 2018 Elsevier Inc. Chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL) are hematopoietic malignancies caused by the constitutive activation of BCR-ABL tyrosine kinase. Although direct BCR-ABL inhibitors, such as imatinib, were initially successful in the treatment of leukemia, many patients developed drug resistance over time due to the gatekeeper mutation of BCR-ABL T315I. In the present study, we found that taxodione, a quinone methide diterpene isolated from Taxodium distichum, significantly induced apoptosis in human myelogenous leukemia-derived K562 cells, which were transformed by BCR-ABL. Taxodione reduced the activities of mitochondrial respiratory chain (MRC) complexes III and V, which appeared to induce the production of reactive oxygen species (ROS). N-acetylcysteine (NAC), an antioxidant agent, canceled taxodione-induced ROS production, reductions in MRC activities, particularly complex V, and apoptotic cell death. Furthermore, in K562 cells treated with taxodione, BCR-ABL and its major signaling molecules, such as STAT5 and Akt were sequestered in mitochondrial fraction, and their localization changes decrease their abilities to stimulate cell proliferation, suggesting that these actions seem to be a mechanism how taxodione functions as an anti-tumor drug. Strikingly, NAC canceled these taxodione-caused anti-cancer effects. Taxodione induced apoptosis in transformed Ba/F3 cells induced not only by BCR-ABL, but also T315I-mutated BCR-ABL through the generation of ROS. Collectively, the present results suggest that in the treatment of leukemia, taxodione has potential as a compound with high efficacy to overcome BCR-ABL T315I mutation-mediated resistance in leukemia cells.

• Access to Document (DOI)

A major component of vitamin E, α-tocopherol inhibits the anti-tumor activity of crizotinib against cells transformed by EML4-ALK

多胡 めぐみ

European Journal of Pharmacology （European Journal of Pharmacology）  825   1 - 9 2018年04月

研究論文（学術雑誌）, 共著, 査読有り,  ISSN  00142999

　概要を見る

© 2018 Elsevier B.V. Crizotinib is an inhibitor of anaplastic lymphoma kinase (ALK) and is of significant therapeutic benefit to patients with non-small cell lung cancer (NSCLC) harboring the EML4-ALK fusion gene. In the present study, we demonstrated that α-tocopherol, a major component of vitamin E, attenuated the effects of crizotinib independently of its anti-oxidant properties. α-Tocopherol significantly inhibited crizotinib-induced apoptosis in cells transformed by EML4-ALK. It also effectively attenuated the crizotinib-induced inhibition of EML4-ALK and its downstream molecules, STAT3 and ERK, and suppressed the inhibitory effects of crizotinib on EML4-ALK-mediated transformation in the focus formation assay. On the other hand, other members of the vitamin E family, namely, β-tocopherol, γ-tocopherol, δ-tocopherol, and α-tocotrienol, and a water-soluble analog of vitamin E, Trolox had no effects on the anti-tumor activity of crizotinib in cells transformed by EML4-ALK. Collectively, these results revealed the risk of the anti-tumor activity of crizotinib being attenuated when it is administrated in combination with vitamin E supplements containing α-tocopherol as a major component.

• Access to Document (DOI)

ARIH2 Ubiquitinates NLRP3 and Negatively Regulates NLRP3 Inflammasome Activation in Macrophages.

多胡 めぐみ

J Immunol. 199 （ 10 ） 3614 - 3622 2017年11月

研究論文（学術雑誌）, 共著, 査読有り

ST2 gene products critically contribute to cellular transformation caused by an oncogenic Ras mutant.

多胡 めぐみ

Heliyon. 3 （ 10 ） e00436 2017年10月

研究論文（学術雑誌）, 共著, 査読有り

Alpha-tocopherol attenuates the anti-tumor activity of crizotinib against cells transformed by NPM-ALK.

多胡 めぐみ

PLoS One. 12 （ 8 ） e0183003 2017年08月

研究論文（学術雑誌）, 共著, 査読有り

Coffee induces vascular endothelial growth factor (VEGF) expression in human neuroblastama SH-SY5Y cells.

多胡 めぐみ

Nutr Neurosci. 20 （ 6 ） 336 - 342 2017年07月

研究論文（学術雑誌）, 共著, 査読有り

Coffee reduces KRAS expression in　Caco-2 human colon carcinoma cells via regulation of miRNAs.

多胡 めぐみ

Oncol Lett. 2017 14 （ 1 ） 1109 - 1114 2017年07月

研究論文（学術雑誌）, 査読有り

Coffee reduces KRAS expression in　Caco-2 human colon carcinoma cells via regulation of miRNAs.

多胡 めぐみ

Oncol Lett. 2017 14 （ 1 ） 1109 - 1114 2017年07月

研究論文（学術雑誌）, 共著, 査読有り

A mouse model reveals that Mfsd2a is critical for unfolded protein response upon exposure to tunicamycin.

多胡 めぐみ

Hum Cell. 30 （ 2 ） 88 - 97 2017年04月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

Coffee extract inhibits adipogenesis in 3T3-L1 preadipocyes by interrupting insulin signaling through the downregulation of IRS1.

多胡 めぐみ

PLoS One 12(3) （ e0173264 ）  2017年03月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

Data on the effect of hypomyelinating leukodystrophy 6 (HLD6)-associated mutations on the TUBB4A properties.

多胡 めぐみ

Data Brief. 11   284 - 289 2017年02月

研究論文（学術雑誌）, 共著, 査読有り

Three Tyrosine Residues in the Erythropoietin Receptor Are Essential for Janus Kinase 2 V617F Mutant-induced Tumorigenesis.

多胡 めぐみ

J Biol Chem. 292 （ 5 ） 1826 - 1846 2017年02月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

Phosphorylated CIS suppresses the Epo or JAK2 V617F mutant-triggered cell proliferation through binding to EpoR.

多胡 めぐみ

Cell Signal. 31   41 - 57 2017年02月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

Roasting Enhances the Anti-Cataract Effect of Coffee Beans: Ameliorating Selenite-Induced Cataracts in Rats.

多胡 めぐみ

Curr Eye Res.    1 - 7 2017年01月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

STAT3 and ERK pathways are involved in cell growth stimulation of the ST2/IL1RL1 promoter.

多胡 めぐみ

FEBS Open Bio. 7 （ 2 ） 293 - 302 2017年01月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

A bis-malonic acid fullerene derivative significantly suppressed IL-33-induced IL-6 expression by inhibiting NF-κB activation.

多胡 めぐみ

Int Immunopharmacol. 40   254 - 264 2016年11月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

Inhibitory effects of flavonoids extracted from Nepalese propolis on the LPS signaling pathway.

多胡 めぐみ

Int Immunopharmacol. 40   550 - 560 2016年11月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

The Extracellular C-loop Domain Plays an Important Role in the Cell Adhesion Function of Aquaporin 0.

多胡 めぐみ

Curr Eye Res.    1 - 8 2016年10月

研究論文（学術雑誌）, 共著, 査読有り

Intracellular NF-HEV/IL-33 harbors essential roles in Ras-induced cellular transformation by contributing to cyclin D1 protein synthesis.

多胡 めぐみ

Cell Signal. 28 （ 8 ） 1025 - 1036 2016年08月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

DDX5 is a novel targeting protein of tumor suppressor ARF

多胡 めぐみ

Seikagaku. 88 （ 2 ） 207 - 210 2016年04月

研究論文（学術雑誌）, 共著

Nepetaefuran and leonotinin isolated from Leonotis nepetaefolia R. Br. potently inhibit the LPS signaling pathway by suppressing the transactivation of NF-κB.

多胡 めぐみ

Int Immunopharmacol. 28 （ 2 ） 967 - 976 2015年10月

研究論文（学術雑誌）, 共著, 査読有り

Hypomyelinating leukodystrophy-associated missense mutation in HSPD1 blunts mitochondrial dynamics.

多胡 めぐみ

Biochem Biophys Res Commun. 462 （ 3 ） 275 - 281 2015年07月

研究論文（学術雑誌）, 共著, 査読有り

Anti-inflammatory activity of flavonoids in Nepalese propolis is attributed to inhibition of the IL-33 signaling pathway.

多胡 めぐみ

Int Immunopharmacol. S1567-5769 （ 15 ） 00019 - 3 2015年01月

研究論文（学術雑誌）, 共著, 査読有り

Arf tumor suppressor disrupts the oncogenic positive feedback loop including c-Myc and DDX5.

多胡　めぐみ

Oncogene. （2015 Jan 15;34(3):314-22.）  34 （ 3 ） 314 - 322 2015年01月

研究論文（学術雑誌）, 査読有り

• Access to Document (DOI)

Coffee inhibits adipocyte differentiation via inactivation of PPARγ.

Aoyagi R, Funakoshi-Tago M, Fujiwara Y, Tamura H. 多胡 めぐみ

Biol Pharm Bull. 37 （ 11 ） 1820 - 1825 2014年09月

研究論文（学術雑誌）

Dexamemthasone suppresses neurosteroid biosynthesis via downregulation of steroidogenic enzyme gene expression in human glioma GI-1 cells.

Koibuchi F, Ritoh N, Aoyagi R, Funakoshi-Tago M, Tamura H*.

Bio Pharm Bull 37(7)   1241-1247 2014年07月

研究論文（学術雑誌）, 共著, 査読有り

Effect of chemical modification on the ability of pyrrolidinium fullerene to induce apoptosis of cells transformed by JAK2 V617F mutant.

Funakoshi-Tago M, Tsukada M, Watanabe T, Mameda Y, Tago K, Ohe T, Nakamura S, Mashino T, Kasahara T.

Int Immunopharmacol 20   258-263 2014年03月

研究論文（学術雑誌）, 共著, 査読有り

Critical role of FANCC in JAK2 V617F mutant-induced resistance to DNA cross-linking drugs.

多胡 めぐみ

Cell Signal. 25 （ 11 ） 2115 - 2124 2013年11月

研究論文（学術雑誌）, 査読有り

Dynamin isoforms decode action potential firing for synaptic vesicle recycling.

多胡　めぐみ

J Biol Chem. 288 （ 26 ） 19050 - 19059 2013年06月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

Critical roles of Myc-ODC axis in the cellular transformation induced by myeloproliferative neoplasm-associated JAK2 V617F mutant.

Funakoshi-Tago M*, Sumi K, Kasahara T, Tago K*.

PLoS One 8(1)   e52844 2013年01月

研究論文（学術雑誌）, 共著, 査読有り

Fullerene derivative prevents cellular transformation induced by JAK2 V617F mutant through inhibiting c-Jun N-terminal kinase pathway.

多胡　めぐみ

Cell Signal. 24 （ 11 ） 2024 - 2034 2012年11月

研究論文（学術雑誌）, 査読有り

• Access to Document (DOI)

Knockdown of proteolipid protein 2 or focal adhesion kinase with an artificial microRNA reduces growth and metastasis of B16BL6 melanoma cells.

多胡　めぐみ

Oncol Lett. 3 （ 1 ） 19 - 24 2012年01月

研究論文（学術雑誌）, 査読有り

Analysis of oncogenic signaling pathway induced by a myeloproliferative neoplasm-associated Janus kinase 2 (JAK2) V617F mutant.

多胡　めぐみ

Yakugaku Zasshi. 132 （ 11 ） 1267 - 1272 2012年

研究論文（学術雑誌）, 単著

Anti-inflammatory activity of structurally related flavonoids, Apigenin, Luteolin and Fisetin.

多胡　めぐみ

Int Immunopharmacol. 11 （ 9 ） 1150 - 1159 2011年09月

研究論文（学術雑誌）, 共著, 査読有り

• Access to Document (DOI)

Aurora kinase A critically contributes to the resistance to anti-cancer drug cisplatin in JAK2 V617F mutant-induced transformed cells.

Sumi K, Tago K, Kasahara T, Funakoshi-Tago M*.

FEBS Lett 585(12)   1884-1890 2011年06月

研究論文（学術雑誌）, 共著, 査読有り

Akt activation through the phosphorylation of erythropoietin receptor at tyrosine 479 is required for myeloproliferative disorder-associated JAK2 V617F mutant-induced cellular transformation.

多胡　めぐみ

Cell Signal. 23 （ 5 ） 849 - 856 2011年05月

研究論文（学術雑誌）, 査読有り

• Access to Document (DOI)

JAK2 is an important signal transducer in IL-33-induced NF-κB activation.

Funakoshi-Tago M*, Tago K, Sato Y, Tominaga S, Kasahara T.

Cell Signal 23(2)   363-370 2011年02月

研究論文（学術雑誌）, 共著, 査読有り

Analysis of the mechanism of polycythemia vera by studying JAK2 mutant-induced signaling pathway.

多胡　めぐみ

Yakugaku Zasshi. 131 （ 8 ） 1183 - 1187 2011年

研究論文（学術雑誌）, 単著, 査読有り

KappaB-Ras is a nuclear-cytoplasmic small GTPase that inhibits NF-kappaB activation through the suppression of transcriptional activation of p65/RelA.

Tago K*, Funakoshi-Tago M, Sakinawa M, Mizuno N, Itoh H.

J Biol Chem 10(7)   30622-30633 2010年10月

研究論文（学術雑誌）, 共著, 査読有り

Licochalcones suppress degranulation by decreasing the intracellular Ca2+ level and tyrosine phosphorylation of ERK in RBL-2H3 cells.

Tanifuji S, Aizu-Yokota E*, Funakoshi-Tago M, Sonoda Y, Inoue H, Kasahara T.

Int Immunopharmacol. 10   769-776 2010年07月

研究論文（学術雑誌）, 共著, 査読有り

The fixed structure of Licochalcone A by alpha, beta-unsaturated ketone is necessary for anti-inflammatory activity through the inhibition of NF-kappaB activation.

Funakoshi-Tago M, Nakamura K, Tsuruya R, Hatanaka M, Mashino T, Sonoda Y, Kasahara T.

Int Immunopharmacol. 10 （ 5 ） 562-571 2010年05月

研究論文（学術雑誌）, 共著, 査読有り

STAT5 activation is critical for the transformation mediated by myeloproliferative disorder-associated JAK2 V617F mutant.

Funakoshi-Tago M, Tago K, Abe M, Sonoda Y, Kasahara T.

J Biol Chem. 285 （ 8 ） 5296-5307 2010年02月

研究論文（学術雑誌）, 共著, 査読有り

Proteolipid protein 2 is associated with melanoma metastasis.

Sonoda Y*,Warita M, Suzuki T, Ozawa H, Fukuda Y, Funakoshi-Tago M, Kasahara T.

Oncol Rep 23 (2)   371-376 2010年02月

研究論文（学術雑誌）, 共著, 査読有り

Pyrrolidinium-type fullerene derivative-induced apoptosis by the generation of reactive oxygen species in HL-60 cells.

Nishizawa C, Hashimoto N, Yokoo S, Funakoshi-Tago M, Kasahara T, Takahashi K, Nakamura S, Mashino T*.

Free Radical Res 43(12)   1240-1247 2009年12月

研究論文（学術雑誌）, 共著, 査読有り

Licochalcone A potently inhibits tumor necrosis factor alpha-induced nuclear factor-kappaB activation through the direct inhibition of IkappaB kinase complex activation.

Funakoshi-Tago M, Tanabe S, Tago K, Itoh H, Mashino T, Sonoda Y, Kasahara T.

Mol Pharmacol. 76 （ 4 ） 745-753 2009年10月

研究論文（学術雑誌）, 共著, 査読有り

The polycythemia vera-associated Jak2 V617F mutant induces tumorigenesis in nude mice.

Abe M, Funakoshi-Tago M*, Tago K, Kamishimoto J, Aizu-Yokota E, Sonoda Y, Kasahara T.

Int Immunopharmacol 9(7-8)   870-877 2009年07月

研究論文（学術雑誌）, 共著, 査読有り

The acute lymphoblastic leukemia-associated Jak2 L611S mutant induces tumorigenesis in nude mice.

Funakoshi-Tago M*, Tago K, Sumi K, Abe M, Aizu-Yokota E, Oshio T, Sonoda Y, Kasahara T.

J Biol Chem 284(19)   12680-12690 2009年05月

研究論文（学術雑誌）, 共著, 査読有り

Licochalcone A significantly suppresses LPS signaling pathway through the inhibition of NF-kappaB p65 phosphorylation at serine 276.

Furusawa J, Funakoshi-Tago M, Tago K, Mashino T, Inoue H, Sonoda Y, Kasahara T.

Cell Signal. 21 （ 5 ） 778-785 2009年05月

研究論文（学術雑誌）, 共著, 査読有り

Glycyrrhiza inflata-derived chalcones, Licochalcone A, Licochalcone B and Licochalcone D, inhibit phosphorylation of NF-kappaB p65 in LPS signaling pathway.

Furusawa J, Funakoshi-Tago M*, Mashino T, Tago K, Inoue H, Sonoda Y, Kasahara T.

Int Immunopharmacol. 9(4)   499-507 2009年04月

研究論文（学術雑誌）, 共著, 査読有り

Dermatophagoides farinae extract induces severe atopic dermatitis in NC/Nga mice, which is effectively suppressed by the administration of tacrolimus ointment.

Oshio T, Sasaki Y, Funakoshi-Tago M, Aizu-Yokota E, Sonoda Y, Matsuoka H, Kasahara T.

Int Immunopharmacol. 9 （ 4 ） 403-411 2009年04月

研究論文（学術雑誌）, 共著, 査読有り

TRAF6 negatively regulates TNFalpha-induced NF-kappaB activation.

Funakoshi-Tago M, Kamada N, Shimizu T, Hashiguchi Y, Tago K, Sonoda Y, Kasahara T.

Cytokine. 45 （ 2 ） 72-79 2009年02月

研究論文（学術雑誌）, 共著, 査読有り

Licochalcone A significantly suppresses LPS signaling pathway through the inhibition of NF-kappaB p65 phosphorylation at serine 276.

Furusawa JI, Funakoshi-Tago M*, Tago K, Mashino T, Inoue H, Sonoda Y, Kasahara T.

Cell Signal 21(5)   778-785 2009年01月

研究論文（学術雑誌）, 共著, 査読有り

Licochalcone A is a potent inhibitor of TEL-Jak2-mediated transformation through the specific inhibition of Stat3 activation.

Funakoshi-Tago M*, Tago K, Nishizawa C, Takahashi K, Mashino T, Iwata S, Inoue H, Sonoda Y, Kasahara T.

Biochem Pharmacol 76(12)   1681-1693 2008年12月

研究論文（学術雑誌）, 共著, 査読有り

Negative regulation of Jak2 by its auto-phosphorylation at tyrosine 913 via the Epo signaling pathway.

Funakoshi-Tago M*, Tago K, Kasahara T, Parganas E, Ihle JN.

Cell Signal 20(11)   1995-2001 2008年11月

研究論文（学術雑誌）, 共著, 査読有り

Mutation of Y925F in focal adhesion kinase (FAK) suppresses melanoma cell proliferation and metastasis.

Kaneda T, Sonoda Y, Ando K, Suzuki T, Sasaki Y, Oshio T, tago M, Kasahara T.

Cancer lett 270   p354-61 2008年11月

研究論文（学術雑誌）, 共著, 査読有り

TRAF6 is a critical signal transducer in IL-33 signaling pathway.

Funakoshi-Tago M*, Tago K, Hayakawa M, Tominaga S-I, Ohshio T, Sonoda Y, Kasahara T.

Cell Signal 20(9)   1679-1686 2008年09月

研究論文（学術雑誌）, 共著, 査読有り

Celecoxib potently inhibits TNFalpha-induced nuclear translocation and activation of NF-kappaB.

Funakoshi-Tago M,Shimizu T,Tago K,Nakamura M,Itoh H,Sonoda Y,kasahara T.

Biochem Pharmacol 76(5)   p662-71 2008年09月

研究論文（学術雑誌）, 共著, 査読有り

Jak2 FERM domain interaction with the erythropoietin receptor regulates Jak2 kinase activity.

Funakoshi-Tago M, Pelletier S, Moritake H, Parganas E, Ihle JN*.

Mol Cell Biol. 28(5)   1792-1801 2008年03月

研究論文（学術雑誌）, 単著, 査読有り

TNF-receptor associated factor 6-deficient fibroblast is sensitive to the TNF-α-induced cell death:Involvement of reactive oxygen species.

Ichikawa D,Funakoshi-Tago M,Aizu-Yokota E,Sonoda Y,Inoue J-I,Kasahara T.

Biochem Biophys Res Commun 351   93-98 2006年12月

研究論文（学術雑誌）, 共著, 査読有り

Two domains of the erythropoietin receptor are sufficient for Jak2 binding/activation and function.

Pelletier S, Gingras S, Funakoshi-Tago M, Howell S, Ihle JN*.

Mol Cell Biol. 26(22)   8527-38 2006年11月

研究論文（学術雑誌）, 単著, 査読有り

Receptor specific downregulation of cytokine signaling by autophosphorylation in the FERM domain of Jak2.

Funakoshi-Tago M, Pelletier S, Matsuda T, Parganas E, Ihle JN*.

EMBO J. 25(20)   4763-72 2006年10月

研究論文（学術雑誌）, 単著, 査読有り

c/EBPalpha inactivation in FAK-overexpressed HL-60 cells impairs cell differentiation.

Hashimoto K, Sonoda Y*, Yamakado M, Funakoshi-Tago M, Yoshida N, Rokudai A, Aizu-Yokota E, Kasahara T.

Cell Signal 18   955-963 2006年07月

研究論文（学術雑誌）, 共著, 査読有り

Functional role of c-Src in IL-1-induced NF-kB activation: c-Src is one of the components of IKK complex.

Funakoshi-Tago M, Tago K, Andoh K, Sonoda Y, Tominaga S-I, Kasahara T*.

Journal of Biochemistry 137   189-197 2005年02月

研究論文（学術雑誌）, 単著, 査読有り

IL-10-induced CCR5 expression in macrophage-like HL-60 cells: Involvement of Erk1/2 and STAT3.

Makuta Y, Sonoda Y, Yamamoto D, Funakoshi-Tago M, Aizu-Yokota E, Takebe Y, Kasahara T*.

Biol Pharm Bull 26 (8)   1076-1081 2003年09月

研究論文（学術雑誌）, 単著, 査読有り

Tumor necrosis factor-induced nuclear factor kappaB activation isimpaired in focal adhesion kinase-deficient fibroblasts.

Funakoshi-Tago M, Sonoda Y, Tanaka S, Hashimoto K, Tago K, Tominaga SI, Kasahara T*.

J Biol Chem 278(31)   29359-29365 2003年08月

研究論文（学術雑誌）, 単著, 査読有り

FAK overexpression upregulates cyclin D3 and enhances cell proliferation via the PKC and PI3-kinase-Akt pathways.

Yamamoto D, Sonoda Y, Hasegawa M, Funakoshi-Tago F, Aizu-Yokota E, Kasahara T.

Cell Signalling 15(6)   575-83 2003年06月

研究論文（学術雑誌）, 単著, 査読有り

TRAF6 and C-SRC induce synergistic AP-1 activation via PI3-kinase-AKT-JNK pathway.

多胡　めぐみ

Eur J Biochem. 270 （ 6 ） 1257 - 1268 2003年03月

研究論文（学術雑誌）

Antiapoptotic action of focal adhesion kinase (FAK) against ionizing radiation.

多胡　めぐみ

Antioxid Redox Signal. 4 （ 3 ） 491 - 499 2002年06月

研究論文（学術雑誌）, 共著

Presence of an inhibitory mechanism on interleukin1a-induced NF-kB activation.

Tago K, Funakoshi M, Mano H, Yanagisawa K, Hayakawa M, Kuroiwa K, Iwahana H, Kasahara T, and Tominaga S-I*,.

Eur J Biochem 268(24)   6526-6633 2001年12月

研究論文（学術雑誌）, 単著, 査読有り

A MEK inhibitor, PD98059 enhances IL-1-induced NF-kB activation by the enhanced and sustained degradation of IkB.

Funakoshi M, Tago K, Sonoda Y,Tominaga S-I, and Kasahara T*.

Biochem Biophys Res Commun 283 (1)   248-253 2001年04月

研究論文（学術雑誌）, 単著, 査読有り

Differential involvement of p38 mitogen-activated protein kinase and phosphatidyl inositol 3-kinase in the IL-1-mediated NF-kB and AP-1 activation.

Funakoshi M, Sonoda Y, Tago K, Tominaga S-I, and Kasahara T*.

Int Immunopharmcol. 1(3)   595-604 2001年03月

研究論文（学術雑誌）, 単著, 査読有り

Suppression of apoptosis by the overexpression of focal adhesion kinases in a human leukemic cell line, HL-60.

Sonoda Y, Matumoto Y, Watanabe S, Funakoshi M, Hanks S K, Kasahara T*.

J Biol Chem 275   16309-16315 2000年11月

研究論文（学術雑誌）, 単著, 査読有り

Anti-apoptotic role of focal adhesion kinase (FAK). Induction of inhibitor-of-apoptosis proteins and apoptosis suppression by the overexpression of FAK in a human leukemic cell line, HL-60.

多胡　めぐみ

J Biol Chem. 275 （ 21 ） 16309 - 16315 2000年05月

研究論文（学術雑誌）

Differential induction of JE/MCP-1 in subclones from a murine macrophage cell line, RAW264.7: Role of kB-3 binding protein.

Ueno M,, Sonoda Y,, Funakoshi M,, Mukaida N,, Nose K,, Kasahara T*.

Cytokine 12(3)   207-219 2000年03月

研究論文（学術雑誌）, 単著, 査読有り