钙离子在顺铂诱导宫颈癌HeLa细胞自噬反应中的作用

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（Ca²⁺）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Ca²⁺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Ca²⁺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。

Role of Ca2+ in cisplatin-induced autophagy of cervical cancer HeLa cells

Objective:To treat the cervical cancer HeLa cells by using calcium inhibitors combined with chemotherapy drugs,and to explore the effect of Ca2+ 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;Ca2+-sensitive fluorescent dye was used to detect the levels of Ca2+ in cytoplasm and mitochondrial;the colocalization of LC3 and P62 were 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 Ca2+-sensitive fluorescent dye detection results showed that compared with cisplatin group,the mitochondrial free Ca2+level and the fluorescence intensity of colocalization of LC3 and P62 protein 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.