Rho激酶抑制剂Y-27632对肝癌HepG2细胞株增殖的影响及机制探讨

Effect and Mechanism of Rho Kinase Inhibitor Y-27632 on Proliferation of HepG2 Cell Line

目的:体外观察Rho激酶抑制剂Y-27632与化疗药物联用对肝癌HepG2细胞增殖能力的影响。方法:Y-27632单独或与顺铂联用处理HepG2细胞株,MTT法检测肝癌HepG2细胞株的增殖能力;流式细胞仪分析HepG2细胞株凋亡情况;Western-blot、RT-PCR检测P53、Bax及Bcl-2蛋白的表达。结果:顺铂、Y-27632分别处理均能显著抑制HepG2细胞的生长,两药联用可产生协同效应(均P<0.05)。顺铂可促进肝癌HepG2细胞株凋亡,Y-27632对细胞凋亡作用不明显,两药联用其凋亡效应显著增强(均P<0.05)。顺铂能显著增加P53和Bax的表达、抑制Bcl-2的表达,Y-27632单用对Bcl-2表达的影响较小,两药联用其效应更明显。单用顺铂或Y-27632均能促进P53、Bax的mRNA表达,并抑制Bcl-2 mRNA表达,两药联用其效应更明显(均P<0.05)。结论:Rho激酶抑制剂Y-27632能显著提高肝癌HepG2细胞株对顺铂的化疗敏感性。

Objective:To explore the effects of Rho kinase inhibitor Y-27632 combined with chemotherapeutic drugs on proliferation of hepatoma HepG2 cells.Methods:The effects of Y-27632 alone or in combination with cisplatin on the proliferation of HepG2 cell line were detected by MTT assay.Flow cytometry was used to analyze the apoptosis of HepG2 cell line.Western blot and RT-PCR were used to detect the protein and mRNA level of P53,Bax and Bcl-2,respectively.Results:Cisplatin and Y-27632 could significantly inhibit the growth of HepG2 cells,and combination of these two drugs could produce synergistic effect(all P<0.05).The results of flow cytometry indicated that cisplatin could promote apoptosis of HepG2 cell line,but Y-27632 had no obvious effect.When the two drugs were combined,the proapoptotic effect was significantly enhanced(all P<0.05).The protein expression of P53 and Bax were increased,and Bcl-2 were decreased by cisplatin,which was more significantly in combination with Y-27632.While,Y-27632 alone had little influence on protein expression of Bcl-2.In addition,the mRNA level of P53 and Bax were elevated,and Bcl-2 was reduced either by cisplatin or Y-27632,which was more obviously in the drug combination group(all P<0.05).Conclusion:Rho kinase inhibitor Y-27632 could improve the chemosensitivity of HepG2 cell line to cisplatin.