Annexin A3抑制顺铂诱导人卵巢上皮癌细胞凋亡

Annexin A3 inhibits apoptosis induced by cisplatin in human epithelial ovarian cancer cells

目的进一步探讨annexin A3能否通过凋亡途径调节卵巢癌细胞对顺铂敏感性。方法运用反转录病毒载体构建及目的细胞感染技术,获得annexin A3稳定上调和稳定下调卵巢癌细胞系,运用annexin V/碘化丙啶分析方法和免疫印迹法检测细胞凋亡率、细胞内caspase蛋白裂解水平和annexin A3表达水平。结果顺铂敏感细胞annexin A3表达水平上调后,60μg/mL顺铂诱导细胞凋亡率由68.72%±1.01%下调至29.13%±2.61%(P<0.05);高浓度顺铂诱导caspase和PARP蛋白裂解。相反,顺铂耐药细胞内annexin A3水平下调后,凋亡率由35.05%±3.06%上升至76.73%±6.42%(P<0.05),低浓度顺铂诱导caspase和PARP蛋白裂解。此外,annexin A3还能够显著抑制卵巢癌细胞内P38 MAPK磷酸化。结论 Annexin A3能通过抑制细胞凋亡调节卵巢癌细胞对顺铂的敏感性。

Objective To determine whether annexin A3 could regulate cisplatin sensitivity through apoptosis induced by cisplatin or not.Methods Retroviral vector constuction and cell infection technology were used to obtain high-level annexin A3 and low-level annexin A3 ovarian cancer cells.Then Annexin V/propidium iodide analysis and Western blot were used to determine the cell apoptosis rate and caspase cleavage.Results When annexin A3 level was up-regualted,the average apoptosis rate induced by 60 μg/mL cisplatin in sensitive cells decreased from 68.72%±1.01% to 29.13%±2.61%（P0.05）,cleavages of caspase and PARP were induced by high level cisplatin.Conversely,in cisplatin resistant cells with low-level annexin A3,the average apoptosis rate was up-reguated from 35.05%±3.06% to 76.73%±6.42%（P0.05）,low level cisplatin stimulated cleavages of caspase and PARP.Besides,annexin A3 may delay phosphorylation of P38 MAPK in ovarian cancer cells.Conclusion All these results show that apoptosis inhibition is a potential mechanism of annexin A3 regulating platinum resistance in ovarian cancer cells.