钙离子在顺铂诱导宫颈癌HeLa细胞自噬反应中的作用

Role of Ca^(2+) in cisplatin-induced autophagy of cervical cancer HeLa cells

目的:采用钙释放抑制剂联合化疗药物作用于宫颈癌HeLa细胞,探讨钙离子(Ca^(2+))在肿瘤细胞自噬反应中的作用,为辅助肿瘤治疗和药物研发提供依据。方法:选择处于对数生长期的人宫颈癌HeLa细胞,随机分为对照组、顺铂组、2-APB组和顺铂联合2-APB组(联合组)。MTT法检测HeLa细胞生存率,钙离子荧光探针法检测HeLa细胞胞浆和线粒体中游离Ca^(2+)水平,共聚焦显微镜检测HeLa细胞中自噬相关蛋白LC3和P62的共定位情况。结果:MTT法检测,与对照组比较,顺铂组和联合组HeLa细胞生存率明显降低(P<0.05);与顺铂组比较,联合组HeLa细胞生存率明显升高(P<0.05)。钙离子荧光探针法检测,与顺铂组比较,联合组细胞株胞浆和线粒体中游离Ca^(2+)水平及LC3和P62蛋白共定位荧光强度明显降低。结论:化疗药物可诱导肿瘤细胞发生自噬,联合钙释放抑制剂给药可以降低肿瘤细胞自噬水平,提高肿瘤细胞对化疗药物的敏感性。

Objective：To treat the cervical cancer HeLa cells by using calcium inhibitors combined with chemotherapy drugs,and to explore the effect of Ca^（2＋）on autophagy in cancer cells,and to provide basis for tumor treatment and drug research.Methods：The human cervical cancer HeLa cells were at logrithmic growth phase randomly divided into control group,cisplatin group,2-APB group,and cisplatin ＋ 2-APB group（combination group）.MTT assay was used to measure the survival rates of HeLa cells;Ca^（2＋）-sensitive fluorescent dye was used to detect the levels of Ca^（2＋）in cytoplasm and mitochondrial;the colocalization of LC3 and P62were examined under confocal microscope.Results：The MTT assay results showed that compared with control group,the survival rates of HeLa cells in cisplatin and combination groups were signifcantly decreased（P〈0.05）.Compared with cisplatin group,the survival rate of HeLa cells in combination group was significantly increased（P〈0.05）.The Ca^（2＋）-sensitive fluorescent dye detection results showed that compared with cisplatin group,the mitochondrial free Ca^（2＋）level and the fluorescence intensity of colocalization of LC3 and P62protein in combination group were significantly decreased.Conclusion：Chemotherapy drugs can induce the autophagy of tumor cells,and calcium inhibitors can increase the sensitity of tumor cells to chemotherapy drugs by reducing the autophagy levels.