丙戊酸钠联合顺铂对HO-8910细胞增殖的影响及机制

Effect of sodium valproate combined with cisplatin on proliferation of HO-8910 cells and the mechanism

目的:探讨丙戊酸钠(VPA)联合顺铂(DDP)对体外培养的人卵巢癌细胞HO-8910细胞增殖抑制和凋亡作用及机制。方法:3 mM浓度的VPA、1μg/m l的DDP及VPA联合DDP分别处理人卵巢癌细胞株HO-8910不同时间(24、48、72 h)后,采用MTT法检测细胞的增殖活性;Hoechst 33258荧光染色观察细胞凋亡形态学改变;RT-PCR检测p53基因mRNA水平表达变化;W estern b lot法检测p53及Stat3蛋白表达变化。结果:VPA对卵巢癌HO-8910细胞株有生长抑制作用,且VPA联合DDP优于单独用药(P<0.05);Hochest 33258荧光染色结果显示,用药后细胞呈现明显的核碎裂、核固缩等典型凋亡改变,VPA联合DDP组细胞凋亡改变较单独用药组明显(P<0.05);RT-PCR检测显示VPA单独用药及联合DDP用药均能提高p53基因的mRNA表达水平;W estern b lot结果显示VPA联合DDP显著增强卵巢癌HO-8910细胞中p53蛋白表达,降低Stat3蛋白的表达,联合用药组效果更明显。结论:VPA单独用药在体外可有效杀伤卵巢癌HO-8910细胞株,与DDP联合应用具有明显的协同作用,推测其可能的作用机制是诱导p53表达水平上调及Stat3表达水平下调。

Objective：To explore the inhibiting effect of sodium valproate combined with cisplatin on proliferation and apoptosis of human ovarian cancer HO-8910 cells in vitro and the mechanism.Methods：Sodium valproate of 3 mM,cisplatin of 1 μg/ml and sodium valproate combined with cisplatin were used to treat human ovarian cancer HO-8910 cells for 24,48 and 72 hours,respectively,then MTT was used to detect the proliferative activity of HO-8910 cells;Hoechst 33258 fluorescence staining was used to observe the morphological changes of cell apoptosis;the change of expression level of p53 gene mRNA was detected by RT-PCR;Western blot was used to detect the changes of expression levels of p53 and stat3 protein.Results：Sodium valproate inhibited the proliferation of human ovarian cancer HO-8910 cells,the clinical efficacy of sodium valproate combined with cisplatin was superior to those of single sodium valproate and single cisplatin（P〈0.05）;the results of Hoechst 33258 fluorescence staining showed that typical apoptotic changes including karyorrhexis and karyopyknosis appeared after treatment,and the apoptotic changes in sodium valproate combined with cisplatin group was more obvious than those in single sodium valproate group and single cisplatin group（P〈0.05）;RT-PCR analysis showed that single sodium valproate and sodium valproate combined with cisplatin both increased the expression level of P53 gene mRNA.Western blot analysis showed that sodium valproate combined with cisplatin increased the expression of P53 protein in human ovarian cancer HO-8910 cells,reduced the expression of Stat3 protein,and the effect was more obvious in sodium valproate combined with cisplatin group.Conclusion：Single usage of sodium valproate can inhibit the proliferation of human ovarian cancer HO-8910 cells in vitro,and sodium valproate combined with cisplatin plays an obvious synergistic effect,it is speculated that the possible mechanism is induction of p53 up-regulation and Stat3 down-regulation.