三肽化合物酪丝缬肽对人肝癌BEL-7402细胞增殖抑制作用的实验研究

Inhibitory mechanism of tyroservaltide on human BEL-7402 hepatocarcinoma cell proliferation

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。

Aim To examine the inhibitory effects of tyroservaltide （YSV） on human hepatocarcinoma BEL- 7402 cells in vitro. Methods＇ An in vitro culture system of hepatocarcinoma BEL-7402 and Chang＇s liver cells was established. We evaluated the effects of YSV on cellular metabolic activity and cytotoxicity against cultured BEL-7402 cells by BrdU incorporation, MTT metabolism, and lactate dehydrogenase （LDH） release. DNA laddering was observed in YSV-treated BEL-7402 cells by agarose electrophoresis. Results After BEL-7402 cells were treated with YSV （ 1 mg · L^-1,0.1 mg · L^-1） for 48 h or 72 h, significant differences were observed between the control and YSV groups. YSV inhibited DNA synthesis （ P 〈 0. 05 ）with a maximum inhibitory rate of 32.53% （ YSV 0. 1 mg · L^-1 for 72 h）. The effects of YSV on the cellular metabolic rate differed in the normal hepatic cell line and the hepatoma cell line. There was an increase in the release of LDH after YSV treatment （ P 〈 0. 05 ）, with a maximal increase of 36. 13%. YSV also induced DNA laddering. When Chang＇s liver cells were treated with YSV, the control and YSV groups differed significantly only when YSV 0. 1 mg · L^-1 was administered for 48 h. Conclusions YSV inhibits the proliferation of BEL-7402 cells, but has almost no effect on Chang＇ s liver cells.