顺铂作用下卵巢癌敏感细胞A2780与耐药细胞A2780/DDP凋亡与自噬水平的变化及意义

Changes and significances of apoptosis and autophagy levels of ovarian cancer sensitive cells-A2780 and resistant cells-A2780/DDP after treated with cisplatin

目的:通过分析自噬对卵巢癌细胞顺铂敏感性的影响,探讨自噬与卵巢癌顺铂耐药性的关系,为卵巢癌治疗提供依据。方法:培养卵巢癌细胞株A2780和A2780/DDP,进行卵巢癌细胞系A2780及A2780/DDP顺铂敏感性的测定,流式细胞仪测定顺铂作用下A2780和A2780/DDP细胞的凋亡率;将10μM顺铂作用48h的A2780细胞和40μM顺铂作用48h的A2780/DDP细胞经处理,采用Western blot法检测Beclin1蛋白的表达;电镜下观察10μM顺铂作用48h的A2780细胞和40μM顺铂作用48h的A2780/DDP细胞经固定染色处理的自噬细胞活性。结果:顺铂主要通过诱导凋亡对卵巢癌耐药细胞A2780/DDP及敏感细胞A2780发挥细胞毒效应;顺铂作用下,耐药细胞的自噬活性明显增强,而敏感细胞自噬活性无明显增强。结论:顺铂诱导耐药卵巢癌细胞发生凋亡的能力较亲代敏感细胞降低,在其发生过程中,反应性自噬活性增强可能与其顺铂耐药性的形成有关,提示对肿瘤细胞的自噬水平进行监测及调控可能为逆转卵巢癌铂耐药提供新的思路。

Objective：To explore the relationship between autophagy and drug resistance to cisplatin in patients with ovarian cancer by analyzing the effect of autophagy on sensitivities of ovarian cancer cells to cisplatin,provide a basis for the treatment of ovarian cancer.Methods：Ovarian cancer cell strains-A2780 and A2780/DDP were cultured,then their sensitivities to cisplatin were detected,flow cytometry was used to detect the apoptosis rates of A2780 cells and A2780/DDP cells after treated with cisplatin;Western blot was used to detect the expression levels of Beclin 1 protein in A2780 cells treated with 10 μM cisplatin for 48 hours and A2780/DDP cells treated with 40μM cisplatin for 48 hours;A2780 cells and A2780/DDP cells were fixed and stained,and the activity of autophagocytes were observed under electron microscope.Results：Cisplatin played cytotoxic effect on ovarian cancer sensitive cells-A2780 and resistant cells-A2780/DDP by inducing apoptosis;after treated with cisplatin,the autophagy activity of sensitive cells increased significantly,while the autophagy activity of resistant cells did not increase obviously.Conclusion：The ability of cisplatin inducing the apoptosis of ovarian cancer resistant cells is lower than that of ovarian cancer parent sensitive cells;the increase of reactive autophagy activity in the course is related to the formation of cisplatin resistance,which indicates that monitoring and regulating the autophagy levels of tumor cells may provide a new idea for reversing cisplatin resistance in ovarian cancer.