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目的观察三肽化合物酪丝缬肽(tyroservaltide,YSV)对体外培养人肝癌BEL7402细胞增殖的抑制作用。方法建立人肝癌BEL7402及Chang氏肝细胞体外培养体系,用BrdU法、MTT法及LDH法,观察YSV对体外培养的BEL7402细胞、Chang氏肝细胞DNA分裂指数、MTT代谢率、LDH释放量的影响,用琼脂糖凝胶电泳观察药物对体外培养BEL7402细胞DNA片段化的影响。结果YSV对BEL7402细胞作用48h、72h时药物浓度为1mg·L-1和0.1mg·L-1组与阴性对照组比较:①能显著抑制肿瘤细胞DNA的合成(P<0.01),抑制率最高为药物浓度1mg·L-1作用72h时可达32.53%,②能表现出明显抑制细胞代谢的作用(P<0.05)其中YSV0.1mg·L-1作用72h时抑制率最高为19.12%,③能增加胞浆内LDH的释放(P<0.05),YSV(1mg·L-1)作用72h时抑制率最高,为36.13%,④能诱导BEL7402肝癌细胞DNA片断化成低分子量DNA,经1%琼脂糖电泳后显示为DNALadder。对正常肝细胞系Chang氏肝仅0.1mg·L-1在48h表现出抑制DNA合成能力的作用。结论YSV抑制人肝癌BEL7402细胞的增殖,对正常肝细胞系Chang氏肝没有影响。
Objective To observe the inhibitory effect of tripeptide tyroservaltide (YSV) on the proliferation of human hepatoma BEL7402 cells in vitro. Methods Human hepatocellular carcinoma cell line BEL7402 and Chang’s hepatocyte culture system were established in vitro. The effects of YSV on DNA fragmentation index, MTT metabolic rate and LDH release in cultured BEL7402 cells and Chang’s hepatocytes were observed by BrdU assay, MTT assay and LDH assay The effect of drugs on DNA fragmentation of BEL7402 cells cultured in vitro was observed by agarose gel electrophoresis. Results The YSV treatment of BEL7402 cells for 48h, 72h drug concentration of 1mg · L-1 and 0.1mg · L-1 group compared with the negative control group: ① can significantly inhibit the synthesis of DNA in tumor cells (P <0.01), the highest inhibition rate (P <0.05). The maximum inhibitory rate of YSV0.1mg · L-1 was 19.12% at 72h when the drug concentration was 1mg · L-1 and 32.53% (P <0.05). When YSV (1 mg · L-1) was applied for 72h, the inhibitory rate was the highest (36.13%). ④ The DNA fragmentation of BEL7402 hepatoma cells was induced to become low molecular weight DNA, Sugar electrophoresis showed as DNALadder. On the normal liver cell line Chang’s liver, only 0.1mg · L-1 at 48h showed an inhibitory effect on DNA synthesis. Conclusion YSV inhibits the proliferation of human hepatocellular carcinoma BEL7402 cells and has no effect on normal liver cell line Chang’s liver.