酪丝缬肽诱导高转移性肝癌细胞HCCLM3凋亡及机制研究  

The Mechanism of Apoptosis Induced by Tyroservatide(YSV) on Highly Metastatic Human Hepatocellular Carcinoma Cell HCCLM3

作　　者：张宁[1] 王鲁[1] 梁英[1] 李薇薇[1] 陈洁[1] 孙惠川[1] 吴伟忠[1] 樊嘉[1] 

机构地区：[1]复旦大学附属中山医院肝癌研究所,上海200032

出　　处：《中国临床医学》2009年第2期203-205,共3页Chinese Journal of Clinical Medicine

基　　金：国家自然科学基金(No30571800);高等学校全国优秀博士论文作者专项基金(No200263)

摘　　要：目的:观察酪丝缬肽(tyroservatide,YSV)诱导高转移性人肝癌细胞HCCLM3的凋亡,分析YSV对肿瘤细胞凋亡相关基因Bax与Bcl-2的影响。方法:建立人肝癌细胞HCCLM3的体外培养体系,用四甲基偶氮唑蓝比色法检测YSV对体外培养的HCCLM3的增殖抑制作用,流式细胞仪检测对照组与YSV剂量组的凋亡率,实时定量PCR法检测YSV对肝癌细胞中Bax与Bcl-2 mRNA表达,Western blotting检测YSV对Bax与Bcl-2蛋白表达的影响。结果:YSV可显著地抑制肝癌细胞HCCLM3的生长,当药物剂量10mg·L-1,作用72h时,抑制率为41.34%(P<0.05)。YSV不同浓度组随着药物作用时间的延长肝癌细胞凋亡率呈升高趋势。实时定量PCR结果显示YSV能上调促凋亡基因Bax mRNA的表达,同时下调凋亡抑制基因Bcl-2 mRNA的表达。Western blotting在蛋白水平上进一步证实了实时定量PCR的结果。结论:YSV可在基因及蛋白水平上调促凋亡基因Bax的表达,同时下调凋亡抑制基因Bcl-2的表达,进而实现其诱导肝癌细胞HCCLM3的凋亡。Objective：To observe the apoptosis effects of highly metastatic potential human hepatocellular carcinoma cell HCCLM3 induced by tripeptide compound tyroservatide （YSV）, and investigate the expression profile of apoptosis-realted gene Bax and BCL-2 in mRNA and protein level affected by YSV. Methods：The culture system of HCCLM3 in vitro was established, the inhibitory effects of YSV on growth of HCCLM3 was detected by MTT assay. The apoptosis rate of the cells in different dose and different time were measured by flow cytometry. Realtime-PCR examined the mRNA expression of Bax and Bcl-2. Western blotting was used to verify the results of realtime-PCR in protein level. Results：YSV could significantly inhibit the growth of HCCLM3, when YSV was used for 72h with the dose of 10mg.L^-1, the inhibitory rate was 41.34%, the difference was significant compared with the control （P〈0.05）. The apoptosis rate of HCCLM3 in different concentration groups of YSV in the same time were higher than that of the control. Realtime-PCR showed that YSV could up-regulated the mRNA expression of apoptosis-induced gene Bax, down-regulated the apoptosis-inhibited gene Bcl-2. Western blotting furthered to verify the outcome of realtime-PCR in protein level. Conclusion：YSV could induce apoptosis of HCCLM3 by up-regulating the expression of Bax, down-regulating the expression of Bcl-2.

关 键 词：酪丝缬肽 凋亡 BAX Bcl-2 

分 类 号：R735.7[医药卫生—肿瘤]