The Effect of HGF on the Growth of Hepatocellular Carcinoma

• Project/Area Number: 15591457
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: Hyogo College of Medicine
• Principal Investigator: OH Koushi Hyogu college of medicine, Faculty of Medicine, Research Assosiate, 医学部, 助手 (80340967)
• Co-Investigator(Kenkyū-buntansha): FUJIMOTO Jirou Hyogo college of medicine, Faculty of Medicine, Professor, 医学部, 教授 (90199373) ; IIMURO Yuuji Hyogo college of medicine, Faculty of Medicine, Assistant Professor, 医学部, 助教授 (30252018) ; KURODA Nobukazu Hyogo college of medicine, Faculty of Medicine, Research Assosiate, 医学部, 助手 (20301658) ; HIRANO Tadamichi Hyogo college of medicine, Faculty of Medicine, Research Assosiate, 医学部, 助手 (90340968) ; OKADA Tosihiro Hyogo college of medicine, Faculty of Medicine, Research Assosiate, 医学部, 助手 (70351799)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥3,300,000 (Direct Cost: ¥3,300,000); Fiscal Year 2004: ¥1,500,000 (Direct Cost: ¥1,500,000); Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
• Keywords: H.C.C / gene herapy / HGF / hepatectomy / liver failure

Research Abstract

Aim : We previously reported that hepatocyte growth factor(HGF) gene therapy ameliorates experimental liver cirrhosis in rats. To elucidate the effect of HGF on hepatocellular carcinoma is crucial in considering the human trials because liver cirrhosis is frequently associated with hepatocellular carcinoma. This study investigated that the effect of HGF on the growth of hepatocellular carcinoma in vivo and in vivo. Methods : 1)We delivered human HGF gene into mice 50ug or rats 500ug by a hydrodynamics-based transfection method that is an in vivo gene transfection procedure by rapid injection of a large volume of naked plasmid DNA solution via the tail vein. Plasma HGF level was measured by ELISA. 2)Two kinds of animal models were used for this study : (a)5x10^6 cells/each tumor of mouse hepatoma cells (Hep1A) were subcutaneously injected into c57BL/6 to establish tumors. HGF gene induction was started when tumor volume was about 100 cubic-millimeters. (b)Wistar Rats were received dieth ylnitrosamine (10 mg/kg/day) to establish liver tumors for 100 days. HGF gene induction was started on day 70 by hydrodynamics-based transfection. These animals were transfected HGF gene by repetitive six times of hydrodynamics-based transfection method and analyzed tumor growth. Results : (1)Plasma HGF levels were extremely high 12 hours after transfection both mice and rats (8.4±3.4,0.6±0.3 ng/ml respectively). (2)No significant difference of tumor growth was seen in mice between HGF transfected mice and non transfected mice. Chemical induced liver tumor volume was similar between HGF transfected rats and non transfected rats by estimation of computed tomography. The biological activities of HGF are produced by the tyrosine phosphorylation of c-Met. The expression of c-Met was observed in liver tumor by Western Blott. We examined the tyrosine phosphorylation of c-Met by Western Blot. The phosphorylation was observed in HGF transducted liver tumors and livers, but it was not observed in non transducted livers. These results indicated that HGF actually acted on liver tumors. Conclusion : These data indicate that HGF does not promote the growth of hepatocellular carcinoma in vivo and in vivo.