Details of a Researcher - TOKINO Takashi

Misc - TOKINO Takashi

Division display 241 - 260 of about 317 ／ All the affair displays >>

Mutations of the beta-catenin gene in endometrial carcinomas

K Kobayashi, S Sagae, Y Nishioka, T Tokino, R Kudo

JAPANESE JOURNAL OF CANCER RESEARCH ( ELSEVIER SCI IRELAND LTD ) 90 ( 1 ) 55 - 59 1999.01

To investigate the contribution of beta-catenin to the development of endometrial carcinoma, we searched for genetic alterations of the beta-catenin gene in primary endometrial carcinomas. Mutational analysis of exon 3 of the beta-catenin gene, encoding the serine/threonine residues for GSK-3 beta phosphorylation, was performed for 35 tumors. Nucleotide sequencing analysis revealed that 5 tumors (5/35, 14%) contained mutations (S33C, S37C, S37F, T41A) that altered potential GSK-3 beta phosphorylation sites. Each of the mutations resulted in the substitution of serine/threonine residues that have been implicated in the down-regulation of beta-catenin through phosphorylation by GSK-3 beta kinase, Furthermore, the incidence of beta-catenin mutations was significantly higher in early-onset (3 of 5) than that in late-onset tumors (2 of 30) (P = 0.014, Fisher's exact test). Replication error (RER)-positive phenotype was not detected in tumors with the beta-catenin gene mutation, although 10 of 35 tumors revealed RER, We performed immunohistochemistry of beta-catenin in 17 cases for which tissue samples were available. We confirmed accumulation of beta-catenin protein in both the nucleus and cytoplasm in 3 tumors, including two in which amino acid alterations had occurred at codon 33 and 37, The other case had no mutation in exon 3, Our results suggested that mutations at serine/threonine residues involved in phosphorylation by GSK-3 beta affected the stability of beta-catenin, Accumulation of mutant beta-catenin could contribute to the development of a subset of endometrial carcinomas, particularly those of the early-onset type.

Molecular cloning, mapping, and characterization of a novel human gene, MTA1-L1, showing homology to a metastasis-associated gene, MTA1

M Futamura, H Nishimori, T Shiratsuchi, S Saji, Y Nakamura, T Tokino

JOURNAL OF HUMAN GENETICS ( SPRINGER-VERLAG TOKYO ) 44 ( 1 ) 52 - 56 1999

Through large-scale sequencing of clones randomly selected from libraries of human cDNAs, we have isolated a novel human gene encoding a product with 59.6% identity in amino acid sequence to human MTA1, a protein associated with tumor invasion and metastasis. This cDNA, named MTA1-L1 (MTA1 like 1), consists of 2736 nucleotides with an open reading frame encoding 668 amino acids. A single 3.0-kb transcript of MTA1-L1 was expressed ubiquitously on Northern blots. Structural analysis of the MTA1-L1 gene revealed 18 exons spanning 8.1 kb of genomic DNA. We assigned the MTA1-L1 locus to chromosomal band 11q12-13.1 by fluorescence in situ hybridization.

DOI PubMed CiNii

Induction of apoptosis in T98G glioblastoma cells by transfection of GML, a p53 target gene

K Ueda, Y Miyoshi, T Tokino, M Watatani, Y Nakamura

ONCOLOGY RESEARCH ( COGNIZANT COMMUNICATION CORP ) 11 ( 3 ) 125 - 132 1999

Expression of the glycosyl-phosphatidylinositol-anchored molecule-like protein (GML) gene, a p53 target, correlates with the sensitivity of some cancer cell lines to anticancer drugs and ionizing radiation. To investigate the function of GML further, we introduced the GML cDNA into various cancer cell lines under control of the tetracycline-regulated system. When we introduced GML into human glioblastoma cell line T98G, which lacks wild-type p53 and expresses no endogenous GML, we observed significant growth suppression accompanied by G(2)/M arrest in two independent, stable cell lines. We confirmed induction of apoptosis by fluorescence-activated cell sorting (FACS) analysis and nuclear staining. Our results indicated that GML could induce apoptosis of T98G without functional p53, and implied that GML plays a crucial role in the apoptotic pathway in some cancer cells.

Significant increase of adenovirus infectivity in glioma cell lines by extracellular domain of hCAR

T Mori, H Arakawa, T Tokino, K Mineura, Y Nakamura

ONCOLOGY RESEARCH ( COGNIZANT COMMUNICATION CORP ) 11 ( 11-12 ) 513 - 521 1999

Recombinant adenoviruses are highly advantageous as vectors for transferring genes into mammalian cells, but the transfer is not efficient in all types of cells. We investigated the effects of four adenoviral receptors [integrin alpha v, integrin beta 3, integrin beta 5, and human coxsackievirus and adenovirus receptor (hCAR)] on adenovirus-mediated transfer of exogenous cDNA into each of 10 glioma cell lines. Transfection efficiency varied widely from one cell line to another (0-100%) when we measured it by infection with AdLacZ, a vector designed to express P-galactosidase. Levels of integrin alpha v and integrin beta 5 expression were similar among the 10 cell lines, but expression of hCAR and integrin beta 3 varied significantly. As these observations indicated a possible correlation between expression of hCAR and the efficiency of gene transfer, we induced the hCAR gene into three glioma cell lines (T98G, U118MG, and U138MG) that expressed hCAR at very low levels and had also revealed low efficiencies of adenoviral gene transfer. In U118MG- and U138MG-derived cells that had regained the ability to express hCAR in stable fashion, adenovirus-mediated gene transfer became highly efficient. Moreover, addition of the peptide corresponding to the extracellular domain of hCAR (ECD-hCAR) by preincubation significantly increased the adenovirus infectivity to these adenovirus-tolerant cells. These results suggest that hCAR could be one of important determinants of the infectivity of adenovirus: and that the ECD-hCAR might be a novel useful tool for improvement of adenovirus-mediated gene therapy against the adenovirus-tolerant cancer cells.

Identification of BAIAP2 (BAI-associated protein 2), a novel human homologue of hamster IRSp53, whose SH3 domain interacts with the cytoplasmic domain of BAI1

K Oda, T Shiratsuchi, H Nishimori, J Inazawa, H Yoshikawa, Y Taketani, Y Nakamura, T Tokino

CYTOGENETICS AND CELL GENETICS ( KARGER ) 84 ( 1-2 ) 75 - 82 1999

BAI1 (brain-specific angiogenesis inhibitor 1) was originally isolated as a p53-target gene specifically expressed in brain. To clarify its function, we have been searching for cellular proteins that associate with the cytoplasmic domain of BAI1. Using its intracellular carboxyl terminus as "bait" in a yeast two-hybrid system, we isolated a cDNA clone named BAIAP2 whose nucleotide sequence would encode a 521-amino acid protein showing significant homology to a 58/53-kDa substrate of insulin-receptor kinase in the hamster. As the expression profile of BAIAP2 examined by Northern blot analysis was almost identical to that of BAI 1, BAIAP2 appears to be active mainly in neurons. In vitro binding assays confirmed that a proline-rich cytoplasmic fragment of BAI1 interacted with the Src homology 3 (SH3) domain of BAIAP2. Double-color immunofluorescent analysis revealed that BAIAP2 was localized at the cytoplasmic membrane when it was coexpressed with BAI1 in COS-7 cells; BAIAP2 not associated with BAI1 was diffused in the cytoplasm. Predominant localization of BAI1 protein in a sub-cellular fraction enriched in growth cones indicated a possible role of BAI1 as a cell adhesion molecule inducing growth cone guidance. As a protein partner of BAI1, BAIAP2 may represent an important link between membrane and cytoskeleton in the process of neuronal growth.

P-2 子宮内膜癌におけるβ-catenin遺伝子の変異

小林寛治, 寒河江悟, 西岡嘉宏, 時野隆至, 工藤隆一

日本産科婦人科學會雜誌 ( 日本産科婦人科学会 ) 51 "S - 360" 1999

CiNii

胃癌におけるβ-カテニンの発現性と遺伝子異常の検索

佐々木泰史, 森本一郎, 伊東文生, 柳原五吉, 今井浩三, 時野隆至

日本癌学会総会記事 58th 1999

J-GLOBAL

Cloning and characterization of BAP3 (BAI-associated protein 3), a C2 domain-containing protein that interacts with BAI1

T Shiratsuchi, K Oda, H Nishimori, M Suzuki, E Takahashi, T Tokino, Y Nakamura

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( ACADEMIC PRESS INC ) 251 ( 1 ) 158 - 165 1998.10

BAI1 (brain-specific angiogenesis inhibitor 1), a p53-target gene specifically expressed in brain, encodes a seven-span transmembrane protein considered to be a member of the secretin receptor family. Using a two-hybrid system, we isolated a cDNA encoding a product that interacts with the cytoplasmic region of BAI1 and designated it BAP3 (BAI1-associated protein 3). The BAP3 product is a novel C2 domain-containing molecule with homology to Munc13 and synaptotagmin. As with Munc13, BAP3 is expressed predominantly in brain. Deletion-mutant analysis revealed that the interaction between BAI1 and BAP3 was not mediated by the Ca domains. Its predominant expression in brain and homology to Munc13 indicate that BAP3, by interacting with BAI1, might be involved in some neuronal function such as regulating release of neurotransmitters. (C) 1998 Academic Press.

DOI PubMed CiNii

Isolation of a novel TP53 target gene from a colon cancer cell line carrying a highly regulated wild-type TP53 expression system

Y Takei, S Ishikawa, T Tokino, T Muto, Y Nakamura

GENES CHROMOSOMES & CANCER ( WILEY-LISS ) 23 ( 1 ) 1 - 9 1998.09

We established a colon cancer cell line SW480-LOWTP53-I carrying a wild-type TP53 transgene that is inducible under control of the lactose operon. Induction of this transgene by isopropyl-beta-D-thiogalactoside (IPTG) arrests growth of the transfected cells. To investigate cellular responses related to the TP53 signaling pathway to induce growth arrest, we applied a differential display method to screen mRNAs isolated from this cell line and looked for genes whose expression was activated or suppressed after induction of wild-type TP53. Subsequent Northern blot analysis confirmed that expression of one novel gene was regulated by wild-type TP53. The cDNA, termed TP53TGI (TP53 target gene I), contained an open reading frame of 270 nucleotides encoding 90 amino acids. Under conditions of cellular stress (ultraviolet irradiation or exposure to bleomycin or cisplatin), expression of TP53TGI was induced in a wild-type TP53-dependent manner, indicating that this gene is likely to play an important role in the signaling pathway of TP53 and may function in response to cellular damage. (C) 1998 Wiley-Liss, Inc.

DOI PubMed CiNii

Cloning and characterization of BAI-associated protein 1: A PDZ domain-containing protein that interacts with BAI1

T Shiratsuchi, M Futamura, K Oda, H Nishimori, Y Nakamura, Takashi Tokino

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( ACADEMIC PRESS INC ) 247 ( 3 ) 597 - 604 1998.06

Brain-specific angiogenesis inhibitor 1 (BAI1), which is a p53-target gene specifically expressed in brain, encodes a seven-span transmembrane protein. Using a two-hybrid system, we isolated a cDNA that encodes a protein, named BAP1 (BAI1-associated protein), which interacts with the cytoplasmic region of BAI1. BAP1 is a novel member of the MAGUK (membrane-associated guanylate kinase homologue) family; it possesses a guanylate kinase domain, WW domains, and multiple PDZ domains. Interaction between BAI1 and BAP1 was mediated by a QTEV motifin the carboxy-terminal region of BAI1 and PDZ domains of BAP1, By immunocytochemical analysis of COS-7 cells transfected with BAI1 and BAP1, both products were co-localized at the cytoplasmic membrane, especially at cell-cell junctions. Cells transfected with BAI1 formed filopodia-like cytoplasmic extensions. These results suggest that BAI1 and BAP1 might be involved in cell adhesion and signal transduction in brain. (C) 1998 Academic Press.

DOI PubMed CiNii

CSR, a scavenger receptor-like protein with a protective role against cellular damage caused by UV irradiation and oxidative stress

HJ Han, Takashi Tokino, Y Nakamura

HUMAN MOLECULAR GENETICS ( OXFORD UNIV PRESS ) 7 ( 6 ) 1039 - 1046 1998.06

Oxidative stress is a pathogenic condition that causes cellular damage and, in a normally functioning cell, several transcription factors respond to this threat by modulating expression of genes whose products ameliorate the altered redox status in some way. We have isolated a novel macrophage scavenger receptorlike gene, CSR (cellular stress response), whose transcription in normal fibroblasts was significantly elevated by exposure to UV radiation or hydrogen peroxide, and pre-treatment with antioxidants prevented induction of CSR. Under conditions of oxidative stress, reactive oxygen species were significantly depleted in CSR-overexpressing cells, indicating that the CSR product protects cells by scavenging oxidative molecules or harmful products of oxidation. Further investigations into the regulation and function of CSR should open a way to understanding the cellular response and the pathogenic processes caused by oxidative stress.

DOI

Cloning and characterization of the murine P2XM receptor gene

G Nawa, T Urano, T Tokino, T Ochi, Y Miyoshi

JOURNAL OF HUMAN GENETICS ( SPRINGER-VERLAG TOKYO ) 43 ( 4 ) 262 - 267 1998

We have isolated the murine counterpart of the human P2XM gene (mP2XM), a P2X purinoceptor that is expressed predominantly in skeletal muscle. The mP2XM gene, consisting of 12 exons that span 10kb of genomic DNA, encodes a 379-amino-acid product with 83% identity to the human homologue. Two potential transmembrane domains (M1 and M2) are present in the predicted product, and a segment resembling the H5 region of voltage-gated ion channels is completely conserved between human and mouse P2XM proteins. Predominant expression of mP2XM in murine skeletal muscle was confirmed by Northern-blot analysis, and a low level of expression was detectable in lung tissue, although human lung does not express P2XM.

DOI PubMed CiNii

Overexpression of GML promotes radiation-induced cell cycle arrest and apoptosis

K Kagawa, T Inoue, T Tokino, Y Nakamura, T Akiyama

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ( ACADEMIC PRESS INC ) 241 ( 2 ) 481 - 485 1997.12

Expression of the GML gene is regulated in a p53-dependent manner and is correlated with the sensitivity of esophageal cancer cells to anti-cancer drugs. To clarify the effect of GML expression on the sensitivity of cancer cells to ionizing radiation treatment, we established cell lines derived from p53-mutant human osteosarcoma HOS and esophageal carcinoma TE10 lines in which GML expression can be induced using the tetracycline-regulable system. Colony formation assay showed that the growth of cells expressing GML are inhibited in response to ionizing radiation, whereas cells not expressing GML were resistant to irradiation. Further investigation demonstrated that GML expression enhances G2/M arrest and apoptosis induced by gamma-irradiation. These results suggest that GML sensitizes cancer cells to ionizing radiation. (C) 1997 Academic Press.

DOI PubMed CiNii

A novel brain-specific p53-target gene, BAI1, containing thrombospondin type 1 repeats inhibits experimental angiogenesis

H Nishimori, T Shiratsuchi, T Urano, Y Kimura, K Kiyono, K Tatsumi, S Yoshida, M Ono, M Kuwano, Y Nakamura, T Tokino

ONCOGENE ( STOCKTON PRESS ) 15 ( 18 ) 2145 - 2150 1997.10

The genetic alteration of p53 is associated with neovascularization during progression of glioma to its more malignant form, glioblastoma, Hence, one or more of the genes transactivated by p53 is likely to function as an angiogenesis inhibitors, We isolated a novel p53-inducible gene that encodes a 1584-amino-acid product containing five thrombospondin type 1 (TSP-type 1) repeats and is specifically expressed in the brain, A recombinant protein corresponding to the TSP-type 1 repeats of this gene product inhibited in vivo neovascularization induced by bFGF in the rat cornea, The expression of this gene, designated BAI1 (brain-specific angiogenesis inhibitor 1) was absent or significantly reduced in eight of nine glioblastoma cell lines, suggesting BAI1 plays a significant role in angiogenesis inhibition, as a mediator of p53.

DOI PubMed CiNii

A novel brain-specific p53-target gene, BAI1, containing thrombospondin type1 repeats inhibits experimental angiogenesis.

T Tokino, H Nishimori, T Shiratsuchi, T Urano, Y Kimura, K Kiyono, K Tatsumi, S Yoshida, M Ono, M Kuwano, Y Nakamura

AMERICAN JOURNAL OF HUMAN GENETICS ( UNIV CHICAGO PRESS ) 61 ( 4 ) A84 - A84 1997.10

Research paper, summary (international conference)

GML sensitizes cancer cells to Taxol by induction of apoptosis

Y Kimura, T Furuhata, T Shiratsuchi, H Nishimori, K Hirata, Y Nakamura, T Tokino

ONCOGENE ( STOCKTON PRESS ) 15 ( 11 ) 1369 - 1374 1997.09

Recently we identified a novel gene, gml, whose expression is regulated in a p53-dependent manner and found that gml expression was correlated with the sensitivity of esophageal cancer cells to anticancer drugs. To further investigate the biological mechanism of gml in determining the chemosensitivity of cancer cells to clinically useful agents, we introduced gml cDNA into TE10, an esophageal cancer cell line that lacks endogenous gml expression. In two resulting stable cell lines which expressed gml cDNA in the absence of wild-type p53, cell death occurred within 6 h after treatment with Taxol. TE10 parent cells or TE10 cells transfected with vector alone displayed relative resistance for 36 h. Induction of gml did not by itself affect viability. Morphological analysis confirmed that the increased chemosensitivity to Taxol conferred by gml was due to apoptosis. These data suggest that reduced expression of gml is likely to be associated with poor response rates to chemotherapy, and that an assay for gml expression might serve a clinical purpose as a predictor of chemotherapeutic sensitivity.

DOI PubMed CiNii

Butyrate activates the WAF1/Cip1 gene promoter through Sp1 sites in a p53-negative human colon cancer cell line

K Nakano, T Mizuno, Y Sowa, T Orita, T Yoshino, Y Okuyama, T Fujita, N OhtaniFujita, Y Matsukawa, T Tokino, H Yamagishi, T Oka, H Nomura, T Sakai

JOURNAL OF BIOLOGICAL CHEMISTRY ( AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC ) 272 ( 35 ) 22199 - 22206 1997.08

Butyrate is a well known colonic luminal short chain fatty acid, which arrests cell growth and induces differentiation in various cell types. We examined the effect of butyrate on the expression of WAF1/Cip1, a potent inhibitor of cyclin-dependent kinases, and its relation to growth arrest in a p53-mutated human colon cancer cell line WiDr, Five millimolar butyrate completely inhibited the growth of WiDr and caused G(1)-phase arrest. WAF1/Cip1 mRNA was rapidly induced within 3 h by treatment with 5.0 mM butyrate, and drastic WAF1/Cip1 protein induction was detected. Using several mutant WAF1/Cip1 promoter fragments, we found that the butyrate responsive elements are two Spl sites at -82 and -69 relative to the transcription start site. We also found that a TATA element at -46 and two overlapping consensus Spl sites at -60 and -55 are essential for the basal promoter activity of WAF1/Cip1. These findings suggest that butyrate arrests the growth of WiDr by activating the WAF1/Cip1 promoter through specific Sp1 sites in a p53-independent fashion.

DOI PubMed CiNii

Cloning of P2XM a novel human P2X receptor gene regulated by p53

T Urano, H Nishimori, HJ Han, T Furuhata, Y Kimura, Y Nakamura, Takashi Tokino

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH ) 57 ( 15 ) 3281 - 3287 1997.08

Through cloning of functional p53-binding sites (p53-tagged sites) from the human genome, we isolated a novel gene inducible by wild-type p53, Its cDNA sequence contained an open reading frame encoding a 431-amino acid peptide that showed a significant homology with members of the P2X family, This protein also revealed a similarity to RP-2, a gene activated in thymocytes undergoing programmed cell death. Northern blot analysis showed that it was expressed predominantly in skeletal muscle, Hence, we designated the gene P2XM (P2X specifically expressed in skeletal muscle), P2XM was localized to chromosomal band 22q11, where frequent loss of heterozygosity has been observed in rhabdoid tumors, Although we detected no genetic alteration in the coding sequences, one of four rhabdomyosarcoma cell lines examined had completely lost expression of this gene, Furthermore, a minor splice variant lacking a part of exon 1 that would encode residues corresponding to transmembrane domain M1 was relatively more abundant in two of seven sarcoma cell lines, one of which was derived from a rhabdomyosarcoma, and the other was derived from an osteosarcoma. The results suggest that P2XM may play a significant role in the proliferation and/or differentiation of skeletal muscle cells and that its altered expression may be involved in the development of some sarcomas.

Genomic structure and chromosomal localization of GML (GPI-anchored molecule-like protein), a gene induced by p53

Y Kimura, T Furuhata, T Urano, K Hirata, Y Nakamura, T Tokino

GENOMICS ( ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS ) 41 ( 3 ) 477 - 480 1997.05

Among its known functions, tumor suppressor gene p53 serves as a transcriptional regulator and mediates various signals through activation of downstream genes, We recently identified a novel gene, GML (glycosylphosphatidylinositol (GPI)-anchored molecule-like protein), whose expression is specifically induced by wildtype p53, To characterize the GML gene further, we determined 35.8 kb of DNA sequence that included a consensus binding sequence for p53 and the entire GML gene. The GML gene consists of four exons, and the p53-binding sequence is present in the 5'-flanking region. in genomic organization this gene resembles genes encoding murine Ly-6 glycoproteins, a human homologue of the Ly-6 family called RIG-E, and CD59; products of these genes, known as GPI-anchored proteins, are variously involved in signal transduction, cell-cell adhesion, and cell-matrix attachment. FISH analysis revealed that the GML gene is located on human chromosome 8q24.3. Genes encoding at least two other GPI-anchored molecules, E48 and RIG-E, are also located in this region. (C) 1997 Academic Press.

DOI PubMed CiNii

Complete NotI restriction map covering the entire long arm of human chromosome 11

F Hosoda, Y Arai, E Kitamura, J Inazawa, M Fukushima, Takashi Tokino, Y Nakamura, C Jones, N Kakazu, T Abe, M Ohki

GENES TO CELLS ( BLACKWELL SCIENCE LTD ) 2 ( 5 ) 345 - 357 1997.05

Background: Human chromosome 11 is one of the autosomes on which many disease genes have been mapped. Many different types of map, including a radiation hybrid map, a genetic map, and an STS-content YAC map, have been constructed for the chromosome, However, a physical map providing accurate physical distances has not yet been established, A chromosome-wide NotI restriction map was constructed to understand the overall feature of the genome organization and to facilitate the positional cloning of disease genes. Results: A complete NotI restriction map of the entire long arm of human chromosome 11 was constructed using linking-clone mapping. This physical map covers 77.6 Mb, from a pericentromeric NotI site to the terminus, and provides the most accurate ordering and distance estimation to date. We also mapped 138 sequence markers in the q13 region that have been poorly mapped previously. Conclusions: The restriction map of the entire long arm of human chromosome 11 is the longest restriction map of the human genome. This mapping has disclosed unique features regarding the organization of the chromosome, indicating that restriction sites of NotI, a CpG-recognition enzyme, are primarily distributed in R (or T) bands and that genetic distance is considerably longer in R (or T) bands than in G bands. The mapping, as well as the dense concentration of mapped markers within the q13 region, should help with positional cloning of the genes associated with various diseases.

DOI PubMed CiNii

　 PREV - NEXT 　

<TOKINO Takashi>

Misc - TOKINO Takashi