胡桃醌联合顺铂对宫颈癌 HeLa 细胞的促凋亡作用及其机制

目的：探讨胡桃醌与顺铂联合应用对宫颈癌HeLa细胞促凋亡作用的机制，阐明其对相关信号转导通路的作用。方法：选取处于对数生长期的HeLa细胞，将其分为对照组、胡桃醌组、顺铂组和胡桃醌＋顺铂组（联合用药组）。采用MTT法观察不同时间点HeLa细胞的增殖抑制率；Hoechst 33258染色法荧光显微镜下观察细胞凋亡形态；Westernblotting法检测各组细胞凋亡相关蛋白Bcl-2、Bax和caspase-3及PI3K/AKt通路中AKt和pAKt表达水平。结果：MTT法，各用药组HeLa细胞增殖于24、48和72h均可被抑制，与对照组比较，胡桃醌组及顺铂组对细胞的抑制较明显，而联合用药组抑制更加显著；与单用药组比较，联合用药组对细胞的抑制更显著。Hoechst 33258染色，胡桃醌组和顺铂组在处理细胞12 h后，出现明显的凋亡细胞形态，联合用药组该现象更明显。Western blotting检测，与对照组比较，12 h后胡桃醌组和顺铂组HeLa细胞中Bcl-2和pAKt蛋白表达水平明显降低，而Bax、Cleave-caspase-3和AKt蛋白表达水平明显升高，联合用药组上述变化更加明显。结论：胡桃醌和顺铂联合应用可通过抑制PI3K/AKt通路，进而促进HeLa细胞凋亡。

Pomotion effect of juglone combined with cisplatin on apoptosis of human cervical cancer HeLa cells and its mechanism

Objective:To explore the mechanism of promotion effect of juglone combined with cisplatin on the apoptosis of human cervical cancer HeLa cells,and to clarify the effects of its associated signal transduction pathways.Methods:The HeLa cells at logarithmic growth phase were divided into control group,juglone group, cisplatin group and juglone combined with cisplatin group (combined treatment group).The inhibitory rates of proliferation of HeLa cells were detected by MTT assay.The apoptosis was detected by Hoechst 33258 staining. The expressions of Bcl-2, Bax and caspase-3, AKt, and pAKt were detected by Western blotting method. Results:The MTT results showed that the HeLa cell proliferation at 24,48 72 h in each drug group was inhibited;compared with control group,the profileration of HeLa cells in juglone group and cisplatin group was significantly inhibited,especially in combined group. Compared with single drug group,the inhibitory effect in combined treatment group was more significantly.After treatment for 12 h,the typical morphological changes of apoptosis were found in juglone group and cisplatin group by Hoechst 33258 staining,especially in combined treatment group. The Western blotting results showed that the expression levels of Bcl-2 and pAKt in HeLa cells in juglone group and cisplatin group 12 h after treatment were decreased obviously,whereas the expression levels of Bax,Caspase-3,and AKt were increased significantly, especially in combined treatment group compared with control group. Conclusion:Juglone combined with cisplatin could inhibit the PI3K/AKt pathway,thereby promoting the apoptpsis of HeLa cells.