APTO-253对紫杉醇或顺铂处理的卵巢癌SKOV3及OVCAR3细胞凋亡及细胞周期的影响
Effects of APTO-253 on apoptosis and cell cycle in SKOV3 and OVCAR3 cells induced by paclitaxel or cisplatin

目的:探讨APTO-253对紫杉醇或顺铂处理的卵巢癌SKOV3及OVCAR3细胞凋亡及细胞周期的影响。方法:①培养SKOV3和OVCAR3细胞,分别加入5 μmol/L APTO-253、4 mg/L顺铂或40 nmol/L紫杉醇、5 μmol/L APTO-253联合4 mg/L顺铂或40 nmol/L紫杉醇,用Caspase-3/7活性方法检测细胞凋亡情况,用Western blot方法测定Cleaved-PARP及Cleaved-Caspase-3蛋白的表达水平。②用5 μmol/L APTO-253处理SKOV3和OVCAR3细胞24 h,采用流式细胞仪检测细胞周期的变化,采用Western blot方法于不同作用时间点(0、1、3、6、12、24 h)测定细胞周期相关蛋白CDK6、cMyc及P21的表达水平。结果:①在SKOV3和OVCAR3细胞中,与APTO-253、紫杉醇或顺铂相比,经APTO-253联合化疗药物处理后细胞凋亡增加(P<0.001),并且Cleaved-PARP及Cleaved-Caspase-3蛋白的表达水平均增加。②在SKOV3和OVCAR3细胞中,APTO-253可导致G1期阻滞、S期和G2/M期细胞减少(P<0.05)。并且随着作用时间的延长,细胞周期相关蛋白CDK6和cMyc表达下降,而P21表达升高。结论:APTO-253能够促进SKOV3及OVCAR3细胞的凋亡,阻滞细胞周期,提高癌细胞对紫杉醇及顺铂的敏感性。
Aim: To investigate the effects of APTO-253 on apoptosis and cell cycle in SKOV3 and OVCAR3 cells induced by paclitaxel or cisplatin.Methods: SKOV3 and OVCAR3 cells were cultured and treated with 5 μmol/L APTO-253,4 mg/L cisplatin or 40 nmol/L paclitaxel,5 μmol/L APTO-253 combined with 4 mg/L cisplatin or 40 nmol/L paclitaxel. The cell apoptosis was detected by Caspase-3/7 activity test and the expressions of Cleaved-PARP and Cleaved-Caspase-3 proteins were measured by Western blot. The cell cycle was analyzed by flow cytometry in SKOV3 and OVCAR3 cells which were induced by 5 μmol/L APTO-253 for 24 h.The expressions of cell cycle associated proteins CDK6, cMyc, and P21 were examined by Western blot following APTO-253 treatment at different time points(0,1,3,6,12,24 h).Results: Compared with the control group, APTO-253 significantly enhanced paclitaxel or cisplatin-induced cell apoptosis(P<0.001)and enhanced the expressions of the Cleaved-PARP and Cleaved-Caspase-3 in both SKOV3 and OVCAR3 cells. APTO-253 treatment led to G1 phase arrest and reduction in S and G2/M phase cells in both SKOV3 and OVCAR3 cells(P<0.05).Cell cycle associated proteins CDK6 and cMyc were downregulated, and P21 was upregulated in a time dependent manner.Conclusion: APTO-253 could not only promote the cell apoptosis and arrest in cell cycle, but also increase the sensitivity of SKOV3 and OVCAR3 cells to cisplatin or paclitaxel