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Biomedical subjects

J Inazawa

Publications and source records attributed to J Inazawa.

At least 91 records · Page 5Linked to original sources
PubMed1998

Aberrations of chromosomes 1 and 17 in six human osteosarcoma cell lines using double-target fluorescence in situ hybridization.

Analysis of six human osteosarcoma cell lines was performed by using double-target fluorescence in situ hybridization (FISH). FISH was applied to interphase nuclei, not to metaphase chromosomes. In this study, numerical aberrations of chromosomes 1 and 17 or structural chromosomal aberrations of chromosome arm 1p or 17p, in which it has been suggested that there are one or more tumor suppressor genes in various malignant tumors, were examined with this technique. All six of the human osteosarcoma cell lines studied had extra copies of chromosomes 1 and 17. A high frequency of deletions (> 60%) in chromosome 1 was found in two cell lines and deletions of chromosome 17 were found in one cell line.

Bone Neoplasms↗
PubMed1998

Analysis of histological therapeutic effect, apoptosis rate and p53 status after combined treatment with radiation, hyperthermia and 5-fluorouracil suppositories for advanced rectal cancers.

The tumour-suppressor gene p53 encodes a transcription factor that plays a critical role in the induction of G1 cell cycle arrest and apoptosis after DNA damage. To clarify the role of the p53 gene and apoptosis in combined hyperthermia, chemotherapy and radiation (hyperthermochemoradiotherapy, HCR therapy) for rectal cancer, we examined the histological response, rate of apoptosis, DNA fragmentation and p53 status in tumours from 28 patients undergoing HCR therapy before surgery and from 22 patients who did not have preoperative treatment. The therapeutic effect of HCR therapy was closely correlated with the rate of apoptosis; the correlation was statistically significant, suggesting that this effect occurs through apoptosis. The incidence of p53 mutations in the treated group were as follows: in tumours resistant to HCR therapy, four of seven (57.1%); intermediately sensitive, 7 of 13 (53.9%); or sensitive, three of eight (37.5%), suggesting that the therapeutic effect and apoptosis rate were related to the p53 status of the tumours to some extent, but the relation was not statistically significant. In the 22 control tumours (non-treated group), the apoptosis rate was 2.0 +/- 1.1%, and there was no significant difference in p53 status compared with the HCR group. Our study indicates that the pathological response to HCR therapy correlates with the rate of apoptosis with statistical significance and that it induces the therapeutic effect more significantly in rectal cancer cells with wild-type p53, although HCR therapy-induced apoptosis also occurs in some rectal cancers with mutated p53. Therefore, this combination therapy can induce an additive or synergistic anti-tumour effect in rectal cancers with wild-type p53 as well as in those with mutated p53 through apoptosis, offering new therapeutic opportunities and a better prognosis.

Antimetabolites, Antineoplastic↗
PubMed1998

ANA, a novel member of Tob/BTG1 family, is expressed in the ventricular zone of the developing central nervous system.

Using a polymerase chain reaction-mediated cloning procedure, we have identified a novel member, termed ANA (from Abundant in Neuroepithelium Area), of Tob/BTG1 family of antiproliferative genes. Molecular cloning and analysis of cDNAs revealed that the human and mouse ANA encoded a protein of 252 amino acids. The amino-terminal half of ANA was homologous to the previously characterized antiproliferative gene products, BTG1, PC3/TIS21/BTG2, and Tob. The human ANA gene was localized at chromosome 21q11.2-q21.1. ANA was expressed in a variety of tissues and cell lines, its expression being high in the ovary, testis, prostate, thymus, and lung. Further analysis revealed that ANA expression was high in the ventricular zone of the developing central nervous system. Finally, overexpression of ANA impaired serum-induced cell cycle progression from the G0/G1 to S phase. In conclusion, ANA is a fourth member of the Tob/BTG1 family that might play roles in neurogenesis in the central nervous system.

Amino Acid Sequence↗
PubMed1998

Grb10/GrbIR as an in vivo substrate of Tec tyrosine kinase.

BACKGROUND: Tec is a member of the recently emerging subfamily among nonreceptor protein-tyrosine kinases (PTKs). Although many members of this family have been shown to be involved in a wide range of cytokine-mediated signalling systems, the molecular mechanism by which they exert in vivo effects remains obscure. To gain insights into the downstream pathways of Tec, we here looked for Tec-interacting proteins (TIPs) by using the yeast two-hybrid screening. RESULTS: One of TIPs turned out to be Grb10/GrbIR, which carries one pleckstrin homology domain and one Src homology 2 domain. Grb10/GrbIR was known to bind receptor PTKs in a ligand-dependent fashion, but not to be phosphorylated on tyrosine residues. In a transient expression system in human kidney 293 cells, however, Grb10/GrbIR becomes profoundly tyrosine-phosphorylated by Tec, but not by Syk, Jak2 or insulin receptor. We also reveal that expression of Grb10/GrbIR suppresses the cytokine-driven and Tec-driven activation of the c-fos promoter. CONCLUSION: Our results indicate a novel role of Grb10/GrbIR as an effector molecule to a subset of nonreceptor PTKs.

Amino Acid Sequence↗
PubMed1998

Amplification on double-minute chromosomes and partial-tandem duplication of the MLL gene in leukemic cells of a patient with acute myelogenous leukemia.

Partial-tandem duplication (PTD) of an internal portion of MLL occurs in some cases of acute myelogenous leukemia (AML) with trisomy 11 or a normal karyotype. This type of MLL rearrangement may be transcribed into an mRNA species that is capable of encoding a partially duplicated protein associated with leukemogenesis. However, although several kinds of oncogenes, especially MYC, are often amplified on double-minute chromosomes (dmins) in hematological malignancies, no amplification of MLL has been reported in AML. Here, we report the first documented case of a patient with AML whose leukemic cells exhibited amplification of MLL on dmins. Furthermore, in this patient, MLL was rearranged in a PTD manner, with in-frame fusion of exons 2 and 6.

Chromosome Aberrations↗
PubMed1997

Detection of Numerical Alterations of Chromosomes 3, 7, 17 and X in Low-grade Intracystic Papillary Tumors of the Breast by Multi-color Fluorescence In Situ Hybridization.

Intracystic papillary tumors of the breast comprise intraductal papillomas and low-grade papillary carcinomas. To determine chromosomal alterations that may be characteristic of these tumors, chromosomes 3, 7, 17 and X were examined in 10 intraductal papillomas and 14 low-grade papillary carcinomas using multicolor fluorescent in situ hybridization. In papillry carcinomas, numerical/structural alterations in chromosome 17 and the accumulation of numerical alterations in chromosomes 3, 7 and X were detected in 21%(3/14) and 15%(2/13) of the cases, respectively. These changes were not detected in papillomas. Loss of chromosome 17p was detected in only one of the 11 DNA-diploid carcinomas, whereas alterations in all four chromosomes were present in a case of DNA-aneuploid carcinoma. Numerical alterations of chromosomes 3, 7, 17 and X do not appear to be involved in the development of low-grade papillary carcinomas with DNA diploidy, but rather accumulate during the formation of DNA-aneuploid carcinoma cells in association with other chromosome alterations.

Journal Article↗
PubMed1997

Molecular cloning and characterization of human keratan sulfate Gal-6-sulfotransferase.

We have previously cloned chondroitin 6-sulfotransferase (C6ST) cDNA from chick embryo chondrocytes. C6ST catalyzes sulfation of chondroitin, keratan sulfate, and sialyl N-acetyllactosamine oligosaccharides. In this study, we report the cloning and characterization of a novel sulfotransferase that catalyzes sulfation of keratan sulfate. This new sulfotransferase cDNA clone was obtained from a human fetal brain library by cross-hybridization with chick C6ST cDNA. The cDNA clone obtained contains a single open reading frame that predicts a type II transmembrane protein composed of 411 amino acid residues. When the cDNA was introduced into a eukaryotic expression vector and transfected in COS-7 cells, keratan sulfate sulfotransferase activity was overexpressed, but C6ST activity was not increased over that of the control. Structural analysis of 35S-labeled glycosaminoglycan, which was formed from keratan sulfate by the reaction with 35S-labeled 3'-phosphoadenosine 5'-phosphosulfate and the recombinant sulfotransferase, showed that keratan sulfate was sulfated at position 6 of Gal residues. On the basis of the acceptor substrate specificity, we propose keratan sulfate Gal-6-sulfotransferase (KSGal6ST) for the name of the newly cloned sulfotransferase. KSGal6ST was assigned to chromosome 11p11. 1-11.2 by fluorescence in situ hybridization. Among various human adult tissues, a 2.8-kilobase message of KSGal6ST was expressed mainly in the brain. When poly(A)+ RNAs from the chick embryo cornea and brain were probed with the human KSGal6ST cDNA in Northern hybridization, a clear band with about 2.8 kilobases was detected. These observations suggest that KSGal6ST may participate in the biosynthesis of keratan sulfate in the brain and cornea.

Amino Acid Sequence↗
PubMed1997

Identification of an Efs isoform that lacks the SH3 domain and chromosomal mapping of human Efs.

Efs was originally found by expression cloning of a mouse embryo cDNA library through its Fyn-SH3 binding capacity (Ishino et al., Oncogene 11, 2331-2338, 1995). Efs has characteristic regions important in intracellular signal transduction; these are an SH3 domain, a cluster of putative ligands for SH2 domains and proline-rich sequences with SH3-binding consensus. In this paper, we report cDNA cloning of human Efs and a variant of it from a hippocampal cDNA library. The human Efs gene was mapped to chromosome 14q11.2-q12 by fluorescence in situ hybridization. We identified two forms of human Efs, designated hEfs1 and hEfs2. hEfs1 represents the human counterpart of original mouse embryo Efs (mEfs1). hEfs2, the newly identified form, is identical to hEfs1, except for its lack of the SH3 domain. hEfs1 and mEfs1 are 80% identical in their amino acid sequences and 100% identical within the SH3 domain. Reverse transcription polymerase chain reaction analysis of adult mouse tissue RNA indicated expression of Efs2 and of Efs1 in various tissues. Evidence suggesting the presence of the Efs2 protein in human tissue was obtained by immunoprecipitation followed by immunoblotting with two different anti-Efs antibodies. Possible functions of Efs2 are discussed.

Adaptor Proteins, Signal Transducing↗
PubMed1997

Ploidy analysis in paraffin-embedded malignant fibrous histiocytoma by DNA cytofluorometry and flourescence in situ hybridization.

To prove the relationship between chromosomal aberration and DNA ploidy in human malignant fibrous histiocytoma (MFH), fluorescence in situ hybridization (FISH) and DNA cytofluorometry were performed in this study. For FISH study, the nucleus of each tumor cell was isolated from paraffin-embedded tissue of nine MFHs. Five chromosome-specific DNA probes (1p36, 1q12, 8q21.3, 11 centromere, and 17 centromere) were hybridized on cell nuclei. Cells with more than three probe signals were regarded as chromosome polysomy. All of the tumors analyzed by FISH had extra copies. The average percentage of polysomy in all tumors was high, ranging from 10.2% to 49.2%. The DNA ploidy patterns, and the percentage of hyperdiploid cells showing a greater DNA content than diploid cells, were obtained from DNA cytofluorometry. Three of nine were diploid patterns and six were non-diploid patterns, and the percentage of hyperdiploid cells in all tumors was high, ranging from 9.1% to 61.9%. The percentage of polysomy could be correlated with the percentage of hyperdiploid cells in each cell. In this study, we found that the DNA ploidy change was closely correlated with aberrations of chromosome copy number in MFH. In addition, the alterations of specific chromosome copy number could be detected in MFH showing diploid cells. Thus, these data indicate that FISH and DNA cytofluorometry are available as a cytogenetic tool for the analysis of interphase nuclei of bone and soft tissue tumors including MFH.

Biopsy↗
PubMed1997

Genomic structure and chromosomal localization of the gene encoding translin, a recombination hotspot binding protein.

Human Translin is a novel DNA end binding protein that specifically recognizes consensus sequences at the breakpoint junctions in chromosomal translocations, mostly involving immunoglobulin (Ig)/T-cell receptor gene segments, in human lymphoid neoplasms. We have recently cloned a full-length human Translin cDNA using peptide sequence information obtained from the purified protein. From comparisons of the amino acid sequences in humans and other vertebrates, we conclude that Translin has been highly conserved during evolution, especially at the leucine zipper motif and the basic region, which is thought to be the DNA binding domain. Analysis of the human and mouse Translin genes revealed that they have identical genomic structures consisting of six exons, five introns, and a GC-rich upstream region. In situ hybridization and physical mapping of somatic cell hybrids allowed localization of the gene to human chromosome 2q21.1.

Amino Acid Sequence↗
PubMed1997

Chromosomal localization of the human and mouse hyaluronan synthase genes.

We have recently identified a new vertebrate gene family encoding putative hyaluronan (HA) synthases. Three highly conserved related genes have been identified, designated HAS1, HAS2, and HAS3 in humans and Has1, Has2, and Has3 in the mouse. All three genes encode predicted plasma membrane proteins with multiple transmembrane domains and approximately 25% amino acid sequence identity to the Streptococcus pyogenes HA synthase, HasA. Furthermore, expression of any one HAS gene in transfected mammalian cells leads to high levels of HA biosynthesis. We now report the chromosomal localization of the three HAS genes in human and in mouse. The genes localized to three different positions within both the human and the mouse genomes. HAS1 was localized to the human chromosome 19q13.3-q13.4 boundary and Has1 to mouse Chr 17.HAS2 was localized to human chromosome 8q24.12 and Has2 to mouse Chr 15. HAS3 was localized to human chromosome 16q22.1 and Has3 to mouse Chr 8. The map position for HAS1 reinforces the recently reported relationship between a small region of human chromosome 19q and proximal mouse chromosome 17. HAS2 mapped outside the predicted critical region delineated for the Langer-Giedion syndrome and can thus be excluded as a candidate gene for this genetic syndrome.

Animals↗
PubMed1997

The brain finger protein gene (ZNF179), a member of the RING finger family, maps within the Smith-Magenis syndrome region at 17p11.2.

Smith-Magenis syndrome (SMS) is caused by a microdeletion of 17p11.2 and comprises developmental and growth delay, facial abnormalities, unusual behavior and sleep problems. This phenotype may be due to haploinsufficiency of several contiguous genes. The human brain finger protein gene (ZNF179), a member of the RING finger protein family, has been isolated and mapped to 17p11.2. FISH analyses of metaphase or interphase chromosomes of 6 patients with SMS show that ZNF179 was deleted in one of the 2 homologs (17p11.2), indicating a possible association of the defect of this gene with the pathogenesis of SMS. Furthermore, using a prophase FISH ordering system, we sublocalized ZNF179 proximally to LLGL which lies on the critical region for SMS.

Abnormalities, Multiple↗
PubMed1997

Cloning of human and mouse cDNAs encoding novel zinc finger proteins expressed in cerebellum and hippocampus.

We identified a novel gene, kf-1, highly expressed in the normal cerebellum but not in the cerebral cortex, the expression of which could have been augmented in the cerebral cortex of a sporadic Alzheimer's disease patient. We cloned human and mouse entire kf-1 cDNAs encoding conserved 79 kDa proteins containing a zinc-binding RING-H2 finger motif at the carboxy-terminus as found in acetylcholine receptor-associated protein (RAPsyn). The 3'-untranslated regions are highly conserved between human and mouse as to constitute a common mRNA secondary structure. In situ hybridization analysis of mouse brain sections revealed strong kf-1 expression in the cerebellum and hippocampus. We propose that KF-1 is involved in membranous protein-sorting apparatus similarly to RAPsyn. We mapped the human kf-1 gene to 2p11.2.

Alzheimer Disease↗
PubMed1997

Frequent association of alternative splicing of NER, a nuclear hormone receptor gene in cancer tissues.

We have detected frequent alternative splicing of a gene that encodes NER, a protein homologous to the retinoic acid receptors, in cancer cells. Western and immunohistochemical analyses disclosed accumulation of a large amount of the aberrant NER product, generated by alternative splicing that caused skipping of an exon corresponding to the DNA-binding domain, in the nucleoli of cells of cancer cell lines and primary cancer tissues. The aberrant protein was detected in 116 of 228 primary cancers developed in various tissues including breast and colon, but was absent in the corresponding normal tissues; it was also detected in 31 of 39 cancer cell lines. This observation may imply that the aberrant NER product has some relation to the development and/or progression of cancers in a variety of human tissues.

Alternative Splicing↗
PubMed1997

cDNA cloning and chromosomal localization of the human telencephalin and its distinctive interaction with lymphocyte function-associated antigen-1.

We have isolated cDNA encoding human telencephalin (TLN), a brain segment-specific neuronal adhesion molecule. Human TLN comprises an NH2-terminal signal peptide, an extracellular region with nine Ig-like domains, a single transmembrane region, and a COOH-terminal cytoplasmic tail. The NH2-terminal five Ig-like domains of TLN were closely related to those of intercellular adhesion molecules (ICAMs)-1 and -3. The TLN gene was mapped to the human chromosome 19p13.2, where the ICAM-1, -3, and -4 (LW) genes are located. Furthermore, we observed lymphocyte function-associated antigen-1 (LFA-1)-mediated adhesion of HL-60 cells on recombinant TLN protein, as well as on ICAM-1. However, the interaction of TLN with LFA-1 on HL-60 cells was divalent cation-independent and phorbol 12-myristate 13-acetate stimulation-independent. We conclude that TLN is a unique neuronal member of ICAM subgroup of the Ig superfamily and propose a novel type of interaction between the Ig superfamily molecule and integrin, which does not require the activation of integrin. TLN on the surface of telencephalic neurons may be a target molecule in the brain for LFA-1-expressing microglia and leukocytes in physiological or pathological conditions.

Amino Acid Sequence↗
PubMed1997

Increased apoptosis rate by hyperthermochemoradiotherapy for advanced rectal cancers.

Apoptosis induced in cancer cells by ionizing radiation, hyperthermia, and 5-fluorouracil (5-FU), termed "hyperthermochemoradiotherapy" (HCR), has been well studied in vitro; however, the role of apoptosis in the tumocidal effect of HCR for primary rectal cancers has not yet been clarified. Therefore, we examined the relationship between the therapeutic effect and induction rate of histological apoptosis in 16 patients with rectal cancers after HCR. Numerous Tunel-positive apoptotic cells were found in the tumor tissue after HCR, but few were found in the tumors which had not received HCR. The histological therapeutic effect was closely correlated to the rate of apoptosis. Thus, we suggest that HCR induces a therapeutic effect mainly through apoptosis in human rectal cancers.

Administration, Rectal↗
PubMed1997

Germline mutation of the RET proto-oncogene in children with total intestinal aganglionosis.

To clarify the pathogenesis of total intestinal aganglionosis, an extremely severe from of neural crest-derived cell migration disorder of the gut, the authors studied possible germline mutations of the RET proto-oncogene (10q11.2) in five pedigrees at high risk for congenital aganglionosis. All five patients analyzed were boys, and one had a family history of Hirschsprung's disease. Genomic DNA was extracted from lymphoblastoid cell lines established from patients and their relatives. Polymerase chain reaction (PCR) products, which were amplified using specific primers (RET; exon 1 approximately 20), were electrophoresed to analyze the single-strand conformational polymorphism (SSCP) patterns. DNA sequences were determined in pedigrees showing abnormal SSCP bands. Among the five patients, three germline mutations were found in the receptor tyrosine kinase domain (exon 15; codons 884, 897, and 904). Amino acid substitutions of the Ret protein were predicted based on the mutated nucleotide changes. Phenotypic variations of congenital aganglionosis may depend on the RET mutation pattern and other genetic or environmental determinants. In our series of patients, male sexuality and germline mutation of the RET tyrosine kinase domain were the most likely factors contributing to this form of Hirschsprung's disease.

Child↗