The Effect of HGF on the Growth of Hepatocellular Carcinoma

HGF对肝细胞癌生长的影响

• 批准号: 15591457
• 负责人: OH Koushi
• 金额: $ 2.11万
• 依托单位: Hyogo College of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591457/
• 关键词: H.C.C; gene herapy; HGF; hepatectomy; liver failure

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 … More 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. Less

目的：我们以前报道肝细胞生长因子（HGF）基因治疗改善大鼠实验性肝硬化。阐明HGF对肝细胞癌的作用在考虑人体试验中至关重要，因为肝硬化经常与肝细胞癌相关。本研究探讨了肝细胞生长因子（HGF）对肝癌生长的影响。研究方法：1）通过尾静脉快速注射大量裸质粒DNA溶液的体内基因转染方法，将人HGF基因导入小鼠50 μ g或大鼠500 μ g。ELISA法测定血浆HGF水平。2)本研究使用两种动物模型：（a）将5 × 10^6个细胞/每个肿瘤的小鼠肝癌细胞（Hep 1A）皮下注射到c57 BL/6中以建立肿瘤。当肿瘤体积约为100 μ m2时开始HGF基因诱导。（B）Wistar大鼠接受饮食 ...更多信息 10 mg/kg/天）以建立肝肿瘤100天。通过基于流体动力学的转染在第70天开始HGF基因诱导。采用基于流体动力学的转染方法，将HGF基因重复转染6次，观察肿瘤生长情况。结果：（1）转染后12 h，小鼠和大鼠血浆HGF水平均极高（分别为8.4± 3.4，0.6 ± 0.3ng/ml）; (2)No HGF转染小鼠与未转染小鼠的肿瘤生长有显著差异。通过计算机断层扫描估计，HGF转染大鼠和未转染大鼠之间的化学诱导的肝肿瘤体积相似。HGF的生物活性是通过c-Met的酪氨酸磷酸化产生的。WesternBlott法检测c-Met在肝癌组织中的表达。我们通过Western Blot检测了c-Met的酪氨酸磷酸化。在HGF转导的肝肿瘤和肝脏中观察到磷酸化，但在未转导的肝脏中未观察到磷酸化。这些结果表明，HGF确实对肝肿瘤有作用。结论：这些数据表明肝细胞生长因子在体内和体内并不促进肝细胞癌的生长。少