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目的:观察健脾益气方含药血清对人MHCC-97H肝癌细胞凋亡的影响,探讨凋亡过程中波形蛋白(Vimentin)和天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)的变化及其意义。方法:采用血清药理学方法,并结合Vimentin裂解剂和Caspase抑制剂,观察7.5%,15%,30%体积浓度健脾益气方含药血清干预对肝癌细胞增殖、侵袭、凋亡,及Vimentin,Caspase-3,聚腺苷二磷酸核糖聚合酶1(PARP-1)蛋白表达的影响。结果:健脾益气方中、高剂量含药血清均可以降低MHCC-97H肝癌细胞中Vimentin蛋白表达(P<0.01),其下调程度与肝癌细胞的增殖抑制率、侵袭抑制率和凋亡率趋势一致;Vimentin裂解组细胞增殖抑制率、侵袭抑制率和凋亡率在各组中均最高;Caspase抑制组细胞的增殖抑制率、侵袭抑制率和凋亡率在各组中均最低。与空白血清组比较,健脾益气方中、高剂量组,Vimentin裂解组Caspase-3蛋白表达上调(P<0.01),Vimentin,PARP-1蛋白表达下调(P<0.01),且PARP-1蛋白出现了裂解片段;Caspase抑制组Caspase-3表达下调(P<0.01),Vimentin,PARP-1蛋白表达无统计学差异,PARP-1无蛋白裂解片段出现。结论:15%的健脾益气方含药血清对人肝癌细胞MHCC-97H的干预效果最佳,可能通过Caspase-3下调细胞中Vimentin蛋白表达,抑制肝癌细胞的增殖与侵袭;同时可能利用Caspase-3/Vimentin形成的正反馈促凋亡信号诱导肝癌细胞发生凋亡。
Objective: To observe the effect of medicated serum of Jianpi Yiqi prescription on the apoptosis of human hepatocarcinoma cell line MHCC-97H and to explore the role of Vimentin and caspase- 3) changes and its significance. Methods: Serum pharmacological methods combined with Vimentin cleavage and Caspase inhibitors were used to observe the proliferation, invasion and apoptosis of the hepatocellular carcinoma cells treated with 7.5%, 15% and 30% , Caspase-3, poly (ADP-ribose) polymerase 1 (PARP-1) protein expression. Results: The middle and high dose of Jianpi Yiqi prescription could reduce the expression of Vimentin protein in MHCC-97H hepatocarcinoma cells (P <0.01), and its down-regulation was related to the proliferation inhibition rate, invasion inhibition rate and apoptosis rate The same trend was observed. The proliferation inhibition rate, invasion inhibition rate and apoptosis rate of Vimentin lysis group were the highest in all groups. The proliferation inhibition rate, invasion inhibition rate and apoptosis rate of Caspase inhibition group were the lowest in all groups. Compared with blank serum group, Caspase-3 protein expression in Vimentin cleavage group was up-regulated (P <0.01), while expression of Vimentin and PARP-1 protein was down-regulated (P <0.01). There was no significant difference in the expression of Vimentin and PARP-1 protein between the Caspase-inhibitor and PARP-1 protein-cleaved fragments. Conclusion: 15% of the drug-containing serum of Jianpi Yiqi prescription has the best intervention effect on human hepatocellular carcinoma cell line MHCC-97H, which may inhibit the proliferation and invasion of hepatocellular carcinoma cells by down-regulating the expression of Vimentin protein in Caspase-3 cells; -3 / Vimentin positive feedback signal to promote apoptosis induced apoptosis in liver cancer cells.