Kazuhiro Hamaoka1,2, Sumiko Nagoshi1,2, Kayoko Sugawara1,2, Kayoko Naiki1,2, Yoshihito Uchida1,2, Mie Inao1,2, Nobuaki Nakayama1,2, Kenji Fujiwara1,2, Satoshi Mochida3,4. 1. Department of Gastroenterology and Hepatology, Saitama Medical University, 38 Morohongo, Moroyama-machi, Iruma-gun, Saitama, 350-0495, Japan. 2. Yokohama Rosai Hospital, Labor Health and Welfare Organization, Yokohama, Japan. 3. Department of Gastroenterology and Hepatology, Saitama Medical University, 38 Morohongo, Moroyama-machi, Iruma-gun, Saitama, 350-0495, Japan. smochida@saitama-med.ac.jp. 4. Yokohama Rosai Hospital, Labor Health and Welfare Organization, Yokohama, Japan. smochida@saitama-med.ac.jp.
Abstract
AIMS: Four single nucleotide polymorphisms (SNPs) exist in the promoter region of the osteopontin (OPN) gene, namely, the SNPs at nucleotide (nt) -155, -616, and -1748 showing linkage disequilibrium to each other, and an independent SNP at nt -443. The significance of these SNPs in the risk of hepatocellular carcinoma (HCC) development was examined in patients with hepatitis C virus (HCV). METHODS: The SNPs at nt -155 and nt -443 were analyzed in 120 patients with HCC. The promoter activity was measured in HepG2 cells by the dual-luciferase reporter assay. The electrophoretic mobility shift assay was performed using nuclear extracts from the cells. RESULTS: Peripheral platelet counts at the time of HCC detection were greater in women with homozygous deletion at nt -155 and C/C or C/T at nt -443 than in those showing other allelic combinations, while no such difference was observed in men. The promoter activity was greater in oligonucleotides with deletions at nt -155 and C at nt -443 than in those with other haplotypes. The mobility shift assay showed double and single complexes with oligonucleotides around nt -155 and nt -443, respectively. Binding activities were greater in deletion than in G in the case of the retarded complex in the former assay and in T than in C in the latter assay. The other complex in the former assay included SRY, showing an equivalent binding activity to oligonucleotides with both alleles. CONCLUSION: OPN promoter SNPs may play a role in the sexual difference in the risk of HCC development through the regulation of OPN expression in patients with HCV.
AIMS: Four single nucleotide polymorphisms (SNPs) exist in the promoter region of the osteopontin (OPN) gene, namely, the SNPs at nucleotide (nt) -155, -616, and -1748 showing linkage disequilibrium to each other, and an independent SNP at nt -443. The significance of these SNPs in the risk of hepatocellular carcinoma (HCC) development was examined in patients with hepatitis C virus (HCV). METHODS: The SNPs at nt -155 and nt -443 were analyzed in 120 patients with HCC. The promoter activity was measured in HepG2 cells by the dual-luciferase reporter assay. The electrophoretic mobility shift assay was performed using nuclear extracts from the cells. RESULTS: Peripheral platelet counts at the time of HCC detection were greater in women with homozygous deletion at nt -155 and C/C or C/T at nt -443 than in those showing other allelic combinations, while no such difference was observed in men. The promoter activity was greater in oligonucleotides with deletions at nt -155 and C at nt -443 than in those with other haplotypes. The mobility shift assay showed double and single complexes with oligonucleotides around nt -155 and nt -443, respectively. Binding activities were greater in deletion than in G in the case of the retarded complex in the former assay and in T than in C in the latter assay. The other complex in the former assay included SRY, showing an equivalent binding activity to oligonucleotides with both alleles. CONCLUSION:OPN promoter SNPs may play a role in the sexual difference in the risk of HCC development through the regulation of OPN expression in patients with HCV.
Entities:
Keywords:
Hepatocellular carcinoma; Osteopontin; SNPs; SRY; Sex difference
Authors: R Kawashima; S Mochida; A Matsui; Y YouLuTuZ; K Ishikawa; K Toshima; F Yamanobe; M Inao; H Ikeda; A Ohno; S Nagoshi; T Uede; K Fujiwara Journal: Biochem Biophys Res Commun Date: 1999-03-24 Impact factor: 3.575
Authors: G N Thalmann; R A Sikes; R E Devoll; J A Kiefer; R Markwalder; I Klima; C M Farach-Carson; U E Studer; L W Chung Journal: Clin Cancer Res Date: 1999-08 Impact factor: 12.531