朱星枚　吴　琳　张　媛　姚　杨　成碧萍　张　丹.西安医学院第一附属医院中心实验室，陕西西安70077；.陕西中医药大学中药化学教研室，陕西西咸新区7046；.第四军医大学生物化学与分子生物学教研室，陕西西安700

细胞自噬与人卵巢癌细胞对顺铂耐药的关系

朱星枚1，2吴琳3张媛3姚杨1成碧萍1张丹2
1.西安医学院第一附属医院中心实验室，陕西西安710077；2.陕西中医药大学中药化学教研室，陕西西咸新区712046；3.第四军医大学生物化学与分子生物学教研室，陕西西安710032

目的探讨细胞自噬与人卵巢癌细胞对顺铂耐药的关系。方法以不同浓度的顺铂（0、1、2、4、8、16 μg/mL）分别作用人卵巢癌细胞A2780及其顺铂耐药株A2780/DDP 48 h，采用MTT法检测卵巢癌细胞的增殖率，透射电镜检测顺铂诱导细胞自噬体的形成，Western blot法测定自噬和凋亡相关蛋白。结果与A2780对照比较，A2780/DDP增殖率明显升高（P＜0.01）；顺铂可以诱导A2780及A2780/DDP中自噬体的形成，且A2780和A2780/DDP中LC3-Ⅱ水平均升高，这种作用可以被自噬抑制剂3-甲基腺嘌呤（3-MA）阻断；顺铂可以诱导A2780中凋亡相关蛋白Cleaved-PARP水平升高，但作用A2780/DDP时影响不大，当3-MA与顺铂同时作用人卵巢癌细胞时，A2780/DDP中的Cleaved-PARP表达水平显著升高（P＜0.01）。结论卵巢癌细胞的耐药可能与顺铂诱导卵巢癌细胞自噬有关，自噬抑制剂3-MA可以抑制顺铂诱导的细胞自噬，同时促进顺铂诱导人卵巢癌细胞凋亡，且增加顺铂耐药株对顺铂的敏感性。

顺铂；耐药；卵巢癌；增殖；细胞自噬；凋亡

［Abstract］Objective To study the relationship between cell autophagy and Cisplatin resistance in the ovarian cancer cells.Methods Cell proliferation was determined by an MTT assay after A2780 and A2780/DDP cells were treated by Cisplatin at different concentrations（0，1，2，4，8，16 μg/mL）for 48 h，the formation of autophagosomes were detected by transmission electron microscopy，both autophagy and apoptosis related protein expression were determined by Western blot.Results Compared with the control of A2780，A2780/DDP proliferation rate was significantly increased（P＜0.01）；Autophagy，characterized by an increase in the number of autophagosomes and LC3-II protein level，was observed in Cisplatin-treated A2780 and Cisplatin-treated A2780/DDP cells，which could be blocked by autophagy inhibitor 3-methyl adenine（3-MA）；Cleaved-PARP protein level was increased in Cisplatin-treated A2780 cells while almost no effect in Cisplatin-treated A2780/DDP cells，however，Cleaved-PARP protein level was significantly increased in Cisplatin and 3-MA-treated A2780/DDP cells（P＜0.01）.Conclusion Resistance of ovarian cancer cells may be associated with autophagy induced by Cisplatin，autophagy inhibitor 3-MA can inhibite autophagy，but promote apoptosis induced by Cisplatin in ovarian cancer cells，and increase the Cisplatin sensitivity to Cisplatin resistant strains.

［Key words］Cisplatin；Resistance；Ovarian Cancer；Proliferation；Autophagy；Apoptosis

卵巢癌是一种常见的妇科恶性肿瘤，严重威胁着妇女生命［1］。临床上常用顺铂治疗卵巢癌，反应率可达70%～80%［2-3］。……