Details of a Researcher - Saya, Hideyuki

Papers - Saya, Hideyuki

Division display 401 - 416 of about 416 ／ All the affair displays >>

Immunocytochemical detection of p53 in human gliomas.

Bruner J. M., Saya H., Moser R. P.

Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 4 ( 5 ) 671 - 674 1991.09

ISSN 0893-3952

Since previous published studies of astrocytomas have shown alterations in the short arm of chromosome 17, and this chromosomal location is that which encodes the p53 protein, we used a monoclonal antibody and immunocytochemistry to detect this protein in a series of brain biopsies. The normal p53 protein has a short half-life and is not detectable using this method. Expression of an altered p53 protein was detected in 29 of 71 brain biopsies, but only in those that showed astrocytic features. p53 expression was detected in 20/32 glioblastomas, 5/12 anaplastic astrocytomas, and 3/5 mixed anaplastic oligo-astrocytomas, but only in astrocytic cells. It could not be detected in any other histologic types of primary brain neoplasms, either benign or malignant. The protein was detected in only 1/11 biopsies interpreted as showing gliosis, but this was in a patient who had previously had a resection for glioblastoma, and may have represented unrecognized infiltrating astrocytoma cells. The p53 protein was also expressed in the nuclei of the two human astrocytoma cell lines examined, U251MG and D54MG. These results are in general agreement with previous detailed chromosomal analyses that have found loss of heterozygosity in up to 60% of malignant astrocytic gliomas.

Detection of loss of heterozygosity in the p53 gene in renal cell carcinoma and bladder cancer using the polymerase chain reaction

Oka K., Ishikawa J., Bruner J. M., Takahashi R., Saya H.

Molecular Carcinogenesis 4 ( 1 ) 10 - 13 1991

ISSN 0899-1987

The human p53 gene, a putative tumor suppressor gene, has a polymorphism in amino acid residue 72. We recently developed a method of detecting codon 72 polymorphism in this gene by digestion of polymerase chain reaction-amplified DNA using an allele-specific restriction endonuclease. This polymorphism allows the identification of loss of heterozygosity for the coding region of the p53 gene in limited tissue samples in a short time without using radioactive materials. We examined 33 patients with renal cell carcinoma and 29 with bladder cancer; heterozygosity in the p53 gene was lost in 60% (6 out of 10 cases) and 73% (8 out of 11 cases) of the renal and bladder tumors, respectively. Additionally, the assay's sensitivity could be improved by using DNA extracted from frozen sections of the tumors. Because the proportions of tumor cells and nontumor cells could be assessed by microscopic evaluation of the frozen sections, we were able to minimize contamination from nontumor cells, which occasionally causes false readings of retained heterozygosity. This simple and sensitive method for detecting loss of heterozygosity in the p53 gene makes it possible to rapidly screen a large number of tissue samples and has the potential to be a useful diagnostic tool for a wide variety of human neoplasms.

Pathways of oncogenesis in primary brain tumors

Steck P. A., Saya H.

Current Opinion in Oncology 3 ( 3 ) 476 - 484 1991

ISSN 1040-8746

Codon 72 polymorphism of the TP53 gene

Ara S., Lee P. S Y, Hansen M. F., Saya H.

Nucleic Acids Research 18 ( 16 ) 4961 1990.08

ISSN 0305-1048

Access to Document (DOI)

Direct sequencing analysis of transmembrane region of human neu gene by polymerase chain reaction

Saya H., Ara S., Lee P. S Y, Ro J., Hung M. C.

Molecular Carcinogenesis 3 ( 4 ) 198 - 201 1990

ISSN 0899-1987

The neu gene in rat neuro/glioblastoma was found to be activated by a single point mutation in the DNA sequence encoding the transmembrane region of the neu-encoded p185 protein. The human homologue of the rat neu gene, termed c-erbB-2 or HER-2, can also be activated in vitro by a similar mutation in the corresponding region. Although the human neu gene was shown to be amplified/overexpressed in a large portion of human breast and ovarian cancer, no reports indicate that the human neu gene is activated by a point mutation in human tumor. To study the possible point mutation of neu gene in human tumors, we characterized the genomic structure in the transmembrane region of human neu gene, which in turn allowed us to determine DNA sequence in this region directly following DNA amplification by polymerase chain reaction. We analyzed 7 tumor cell lines (2 breast cancer, 1 neuroblastoma, 1 rhabdomyosarcoma, and 3 glioma) and 11 tumor tissue samples (8 breast and 3 ovarian cancers). No mutation was found in the transmembrane region of human neu gene. Our results suggest that unlike the rat neuro/glioblastoma, the single point mutation in the transmembrane region of the human neu gene is a rare event in human tumors. In this study, we developed a technique for direct DNA sequencing of the transmembrane region of the human neu gene. This technique makes it possible to screen a large number of tumor samples.

Modification of a commercially available method of extracting plasmid DNA

Lee P. S Y, Ara S., Saya H.

BioTechniques 8 ( 4 ) 386 1990

ISSN 0736-6205

Application of polymerase chain reaction for rapid subcloning of cDNA inserts from λgt11 clones

Herrmann J., Lee P., Saya H., Nakajima M.

BioTechniques 8 ( 4 ) 376 - 381 1990

ISSN 0736-6205

Cell kinetic analysis of human brain tumors by bivariate flowcytometric measurement of cellular DNA content and amount of incorporated bromodeoxyuridine

Okuda Y., Taomoto K., Saya H., Ijichi A., Kudo H., Kokunai T., Tamaki N., Matsumoto S.

Brain and Nerve 41 ( 4 ) 383 - 390 1989

ISSN 0006-8969

Antigen common to several species, recognized by a rat monoclonal antibody raised against syngeneic rat bladder tumor

Eto H., Saya H., Nakata M., Mizoguchi A., Kamidono S.

International Journal of Cancer 44 ( 3 ) 454 - 459 1989

ISSN 0020-7136

A rat monoclonal antibody (MAb) termed RS-II (Ig M) was obtained by syngeneic immunization with rat bladder tumor cells induced by N-butyl, N-hydroxybutylnitrosamine (BBN). Immunocytochemical analysis showed that RS-II is also reactive with mouse, dog and human bladder tumor-cell lines and some other human tumor-cell lines but not myeloma or leukemia cells. Immunohistochemical examination of paraffin-embedded tissues has shown that RS-II is reactive with mouse, rat, dog and human bladder tumors (5/5, 5/5, 1/1, 31/49) and some other tumors, but not with normal human urothelium or normal rat tissues. The antigen is expressed on the majority of low-grade or well-differentiated tumors, but less on advanced, invasive tumors. The immunofluorescence staining pattern of cultured cells was cytoplasmic and filamentous. Immunoblotting revealed that the antigen is a M(r) 54,000 and M(r) 56,000 protein which was extracted with difficulty from the cultured cells with Triton X-100. The antigen was also detected in the culture supernatant by means of ELISA. Our results suggest that the epitope is expressed on cytoskeletons common to a wide variety of mammalian cells at a certain stage of differentiation.

Neuronal cell differentiation of human neuroblastoma cells by retinoic acid plus herbimycin A

Preis P. N., Saya H., Nadasdi L., Hochlaus G., Levin V., Sadee W.

Cancer Research 48 ( 22 ) 6530 - 6534 1988

ISSN 0008-5472

We investigated the effect of retinoic acid (RA) and herbimycin A (herb-A) on cell growth, cell differentiation, and colony formation of human neuroblastoma cell lines. The neuroblastoma line SK-N-SH expressed both neuroblast and nonneuronal phenotypes, whereas its subclone SH-SY5Y and the Kelly cell line were predominantly neuroblastic. Both herb-A and RA, given alone, moderately reduced cell growth and colony formation of the neuroblastic cell lines. Growth curve analyses with SK-N-SH suggested that herb-A greatly reduced the number of initially growing cells, whereas RA slightly enhanced initial cell growth. Morphological changes were determined with the use of rhodamine-phalloidin staining of actin. Retinoic acid caused an increase in the fraction of neuroblast cell in SK-N-SH, and conversely of nonneuronal cells in SH-SY5Y and Kelly cell lines. Both drugs also caused partial differentiation towards a neuronal phenotype, and herb-A induced selective lysis of nonneuronal cells of SK-N-SH. Because of their discrepant effects, RA (10 μM) and herb-A (236 nM) were tested in combination at a concentration that had only moderate effects when given alone. The combination further reduced cell growth and colony formation and dramatically enhanced differentiation towards a neuronal morphology. The Kelly cell line with amplified N-myc genome, which correlates with clinical progression of neuroblastoma, was also sensitive to RA plus herb-A. These results recommend the combination of RA and herb-A for differentiation therapy of neuroblastoma.

A hypothesis for myeloschisis

Oi S., Saya H., Matsumoto S.

Journal of Neurosurgery 68 ( 6 ) 947 - 954 1988

ISSN 0022-3085

A hypothesis for embryopathogenesis of myeloschisis is described on the basis of experimental studies analyzing the stage specificity and immunohistochemical/histological characteristics of the exposed neural tissue (placode). Myeloschisis developed in six fetuses among 205 chick embryos treated in various stages with teratogens including ethylnitrosourea, and anticonvulsant and antipyretic agents. All but one case (with associated cephalothoracopagus) demonstrated myeloschisis in the thoracic region with a lamina defect of two and three levels. No fetus was exposed to a teratogen prior to or within Hamburger and Hamilton stage 12 (45 to 49 hours postincubation), when the neuropore closes. Immunohistochemical studies of chick myeloschisis clearly indicated that neuron-specific enolase-positive elements were extremely active only in the overgrown placode, corresponding to the histological findings with Kluver-Barrerra's special stain. These findings were compared with observations in a case of myeloschisis in a human neonate. The results of this study imply the possibility of another mechanism for the embryopathogenesis of myeloschisis: namely, the overgrowth and reopening hypothesis.

Pineoblastoma with neuronal differentiation. Immunohistochemical and immunocytochemical studies

Okuda Y., Taomoto K., Saya H., Ijichi A., Kokunai T., Tamaki N., Matsumoto S.

Journal of Neuro-Oncology 6 ( 2 ) 193 - 198 1988

ISSN 0167-594X

Protein-tyrosine kinase activity and pp60(v-src) expression in whole cells measured by flow cytometry

Preis P. N., Waldman F. M., Frackelton A. R., Saya H., Levin V. A.

Cancer Research 48 ( 16 ) 4633 - 4638 1988

ISSN 0008-5472

The expression and phosphotyrosine activity of pp60(v-src) were measured in the B31 avian sarcoma virus-transformed rat cell line by flow cytometry using monoclonal antibodies against pp60(v-src) (EB7) and phosphotyrosine (1G2). Although the immunocytochemical staining was markedly heterogeneous, binding of both antibodies was significantly greater to B31 cells than to untransformed Rat 1 cells. Binding of 1G2 to phosphotyrosine residues was specific; it was entirely inhibited by adding excess phenylphosphate but was not affected by phosphoserine or phosphothreonine. The relationship between the amount of phosphorylated tyrosine measured by phosphoamino acid analysis was linear in vanadate-treated BALB/c 3T3 cells. Treatment of B31 cells for 48 h with herbimycin A, a benzenoid ansamycin antibiotic, to decrease the expression and tyrosine kinase activity of pp60(v-src) caused reductions of 42% in anti-pp60(v-src) and 58% in anti-phosphotyrosine antibody immunofluorescence. DNA staining with the fluorescent dye propidium iodide showed no cell cycle specificity in the binding of either antibody. Herbimycin A also caused the transformed cell line to revert to the morphology, actin configuration, and growth behavior of untransformed cells; these changes were reversed within 12 h after removal of the drug. Flow cytometric evaluation of tyrosine kinase expression and activity was fast and easy, and the results correlated well with other measures of cell phenotype. This technique can be used to quantitate the effects of drugs on oncogenic proteins such as pp60(v-src) and their associated tyrosine kinase activity.

Detection of human glioma-associated antigen by rat monoclonal anibody raised against syngeneic rat glioma cells

Saya H., Masuko T., Kokunai T., Yagita H., Ijichi A., Taomoto K., Tamaki N., Matsumoto S., Hashimoto Y.

Journal of Neurosurgery 65 ( 4 ) 495 - 502 1986

ISSN 0022-3085

A monoclonal antibody termed 'FR77' was obtained from a hybridoma clone established by fusion between P3x63Ag8.653 mouse myeloma cells and spleen cells of a Fischer F344 rat hyperimmune to syngeneic 9L/R₃ glioma cells. Immunoperoxidase staining of various cultured cells showed that FR77 was reactive to both rat and human glioma cells, but was not reactive with other nonglioma cells. Immunohistochemical examination of paraffin-embedded or cryostat-frozen sections of various human tissues revealed that FR77 was strongly reactive with glioblastoma, grade III astrocytoma, and craniopharyngioma; partially reactive with intracerebral primitive neuroectodermal tumor, pineoblastoma, and desmoplastic medulloblastoma; and weakly reactive with low-grade astrocytoma. It was not reactive with other types of brain tumors and normal human tissues tested. The FR77-defined antigen was observed to be predominantly localized in the cytoplasm of antigen-bearing cells as suggested by the immunostaining pattern, but part of it was also expressed on the cell surface of glioma cells as demonstrated by a complement-mediated cytotoxic test. Fractionation of the antigenic component and periodic acid treatment of tumor tissue bearing the FR77-defined antigen indicated that the antigen is of a neutral glycolipid nature and that the antigenic determinant to FR77 is present on its sugar portion.

Derivation of a brain tumor-selective monoclonal antibody from hybridoma between mouse myeloma and rat spleen cells immune to syngeneic glioma

Saya Hideyuki, Masuko Takashi, Hashimoto Yoshiyuki

Japanese Journal of Cancer Research 76 ( 12 ) 1198 - 1202 1985.12

ISSN 0910-5050

Fischer 344 (F344) rats hyperimmunized with syngeneic 9L R₈ glioma cells produced antibody selective to glioma cells. Hybridomas prepared from the spleen cells of the immunized rat were cloned, and we obtained a hybridoma clone which produced monoclonal IgM antibody, termed FR77, that showed selectivity to glioma cells. Immunoperoxidase staining of cultured cells revealed that FR77 was reactive with 3 lines of rat glioma cells but not with normal F344 rat fibroblasts. Immuno-histochemical staining of F344 rat tissue sections with biotinylated FR77 demonstrated that FR77 could bind with glioma tissue developed by intracerebral injection of 9L R₈ glioma cells but not to normal parts of the brain tissues and other normal tissues tested. The FR77-defined antigen was observed to be mainly localized in cytoplasm of glioma cells but a small portion of the antigen was also detected on the glioma cell surface.

Angiographic classification of aneurysms of the horizontal segment of the anterior cerebral artery

Wakabayashi Toshimitsu, Tamaki Norihiko, Yamashita Hideyuki, Saya Hideyuki, Suyama Tohru, Matsumoto Satoshi

Surgical Neurology 24 ( 1 ) 31 - 34 1985

ISSN 0090-3019

Ten aneurysms of the horizontal portion of the anterior cerebral artery (A₁) were analyzed. These 10 aneurysms were classified into three types according to the mode of the origin of the neck of the aneurysm from the A₁ segment: (a) an aneurysm originating from the junction of the A₁ segment and the lenticulostriate artery, (b) one from the proximal end of the fenestration of the A₁ and (c) one from the top of the A₁ loop (elongation and kinking of the A₁ segment). In these aneurysms, their size, the side of the A₁ segment with an aneurysm, the multiplicity of aneurysms, the coexistence of hypertension, the grade of the patients, the laterality of the subarachnoid blood clots (as seen on computed tomography scans) and surgical outcome were analyzed.

Access to Document (DOI)

　 Previous page - Next page 　