细胞自噬及人卵巢癌细胞对顺铂耐药关系.docVIP

细胞自噬及人卵巢癌细胞对顺铂耐药关系 　　[摘要] 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 mu;g/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 lt; 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 lt; 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。　　[关键词] 顺铂；耐药；卵巢癌；增殖；细胞自噬；凋亡　　[中图分类号] R965 [文献标识码] A [文章编号] 1674-4721（2016）09（a）-0012-05　　[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 mu;g/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 lt; 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 lt; 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