哌立福辛联合顺铂对卵巢透明细胞癌ES2细胞系凋亡及Caspase-3和PARP表达的影响

发布时间：2018-06-24 03:05

本文选题：哌立福辛 + 顺铂　；参考：《现代妇产科进展》2015年10期

【摘要】：目的:探讨哌立福辛(perifosine)与顺铂(cisplatin)联合对人卵巢透明细胞癌ES2细胞系凋亡及Caspase-3和PARP表达的影响。方法:以不同浓度(0、2、4、6、8和10μmol/L)哌立福辛作用于人卵巢透明细胞癌ES2细胞系48h。CCK-8法检测细胞增殖抑制情况。以哌立福辛最小有作用剂量与不同浓度(0、10、40、70和100μmol/L)顺铂联合处理细胞48h,以金氏公式筛选出协同抑制作用最明显的联合剂量,以该联合剂量处理ES2细胞48h后,DAPI染色法、流式细胞仪检测细胞凋亡情况;用Giemsa染色检测细胞有丝分裂指数;Western blot法检测细胞中Caspase-3(cleaved)及PARP(cleaved)蛋白的表达情况。结果:CCK-8法检测提示,哌立福辛作用48h时对ES2细胞的最小作用剂量为4μmol/L。金氏公式计算结果提示,4μmol/L哌立福辛+40μmol/L顺铂联合用药协同抑制细胞增殖作用最强(q=1.21);与哌立福辛或顺铂单独作用相比,DAPI染色发现联合用药组镜下可见大量的核碎裂及凋亡小体;流式细胞仪检测发现,联合用药能协同增加细胞凋亡率(q=1.32,P0.01)。Giemsa染色结果显示,联合用药能协同抑制细胞有丝分裂(P0.01)。Western blot检测发现,联合用药能明显增加细胞中Caspase-3(cleaved)蛋白的表达,降低PARP(cleaved)蛋白的表达。结论:哌立福辛与顺铂联合能协同通过增加Caspase-3(cleaved)蛋白表达及降低PARP(cleaved)蛋白表达,促进人卵巢透明细胞癌ES2细胞发生凋亡。
[Abstract]:Aim: to investigate the effects of pirifampicin (perifosine) combined with cisplatin (cisplatin) on apoptosis and expression of Caspase-3 and PARP in human ovarian clear cell carcinoma cell line ES2. Methods: the proliferation inhibition of human ovarian clear cell carcinoma ES2 cell line was detected by different concentrations of pirifoxine (0 渭 mol / L and 10 渭 mol / L) by CCK-8 method. The cells were treated with pirifoxine at a minimum active dose of 4070 and 100 渭 mol / L cisplatin for 48 h. The most obvious synergistic inhibitory dose was screened by Kim's formula. The cells were treated with the combined dose for 48 h and then treated with DAPI staining method. The cell apoptosis was detected by flow cytometry and the expression of Caspase-3 (cleaved) and PARP (cleaved) protein was detected by Giemsa staining and Western blot assay. Results the minimum dose of pirifoxine on ES2 cells was 4 渭 mol / L after 48h exposure. The results of Kim's formula showed that 4 渭 mol / L pirifoxine 40 渭 mol / L cisplatin combined with cisplatin had the strongest synergistic inhibitory effect on cell proliferation (q1. 21), compared with pirifampicin or cisplatin alone, a large number of nuclear fragmentation and apoptotic corpuscles were found in the combined treatment group. The results of flow cytometry showed that the combined drug could increase the cell apoptosis rate (QN 1.32 P0.01). Giemsa staining results showed that the combined drug could inhibit the cell mitosis (P0.01). Western blot assay showed that the combined treatment could significantly increase the expression of Caspase-3 (cleaved) protein in the cells. The expression of PARP (cleaved) protein was decreased. Conclusion: the combination of pirifampicin and cisplatin can promote apoptosis of human ovarian clear cell carcinoma ES2 cells by increasing the expression of Caspase-3 (cleaved) protein and decreasing the expression of (cleaved) protein.
【作者单位】：中国医学科学院北京协和医学院北京协和医院妇产科;
【分类号】：R737.31

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