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作 者:张盛周[1] 张宏霞[1] 潘飞燕[1] 李朝军[1]
机构地区:[1]南京师范大学生命科学学院江苏省分子医学生物技术重点实验室
出 处:《肿瘤防治研究》2008年第5期309-312,320,共5页Cancer Research on Prevention and Treatment
基 金:安徽高校省级自然科学研究重点资助项目(KJ2007A119ZC)
摘 要:目的探讨重组腺病毒介导的siRNA技术下调c-Met表达对c-Met过表达肝癌细胞在体外生长和成瘤性的影响。方法用重组腺病毒介导的siRNA技术下调c-Met的表达;通过Western blot检测蛋白质的表达水平;以MTT法和细胞计数法检测细胞的生长和增殖情况;以流式细胞术检测细胞周期变化情况;以软琼脂克隆形成实验考察细胞的克隆形成能力。结果重组腺病毒介导的siRNA可使HCCLM3细胞c-Met的表达量下调90%以上。下调c-Met表达可使HCCLM3细胞生长停留在G1/G0期,增殖速率下降30%以上;下调c-Met表达可使HCCLM3细胞克隆形成能力下降约78%(P<0.01)。结论重组腺病毒介导的siRNA下调c-Met表达具潜在的肝癌基因治疗价值。Objective To investigate the effects of knockdown of c-Met by adenovirus-delivered siRNA on the growth of hepatocellular carcinoma cells in vitro. Methods c-Met was knocked down by adenovirus- delivered siRNA. The expression of protein was detected by Western blot. Cell proliferation was determined by MTT assay and Cell count experiments. Cell cycle was analyzed by flow cytometry. Cell colony formation ability was assessed by soft agar colony formation assay. Results The adenovirus-delivered siRNA could decrease the c-Met expression of HCCLM3 cells by over 90%. Knockdown of c-Met could arrest HCCLM3 cells at G1/G0 and inhibit the proliferation of HCCLM3 cells by over 30% (P〈0. 01). Furthermore, Knockdown of c-Met could reduce HCCLM3 cells colony formation by 78% (P〈0. 01), Conclusion Knockdown of c-Met expression by adenovirus-delivered siRNA may be a potential approach for hepatocellular carcinoma gene therapy.
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