细胞自噬与卵巢癌SKOV3细胞及其耐药株SKOV3/DDP对顺铂敏感性的关系  

The relationship between autophagy and the sensitivity to cisplatin in ovarian cancer SKOV3 and SKOV3/DDP cells

作　　者：吴小华 陈亚楠[1] 杨波[1] 周楠[1] 王娜[1] 许鹏宇[1] 

机构地区：[1]中国人民解放军白求恩国际和平医院妇产科,河北石家庄050082

出　　处：《肿瘤》2013年第10期862-867,共6页Tumor

摘　　要：目的:探讨细胞自噬与卵巢癌SKOV3细胞及其顺铂(cisplatin,DDP)耐药株SKOV3/DDP对DDP敏感性的关系。方法:在透射电子显微镜下观察SKOV3和SKOV3/DDP细胞的自噬体形成情况;MTT法筛选自噬抑制剂3-甲基腺嘌呤(3-methyladenine,3-MA)对SKOV3和SKOV3/DDP细胞的无毒剂量,并检测3-MA联合DDP干预后SKOV3和SKOV3/DDP细胞对DDP敏感性的变化;分别采用FCM法和蛋白质印迹法检测单用DDP和3-MA及DDP联合3-MA作用后,SKOV3和SKOV3/DDP细胞的凋亡率和自噬标志蛋白微管结合蛋白1轻链3(microtubule-associated protein 1 light chain 3,MAP1LC3,LC3)和自噬相关蛋白Beclin1的表达。结果:透射电子显微镜下观察可见SKOV3/DDP细胞内有多个自噬体形成,SKOV3细胞内仅可见少量自噬体形成。与DDP单独作用相比,DDP联合3-MA作用时DDP对SKOV3/DDP细胞的半数抑制浓度(half inhibitory concentration,IC50)值由(17.71±0.15)μg/mL下降到(8.85±0.11)μg/mL(P<0.05),细胞凋亡率由(6.93±0.94)%提高到(14.82±0.85)%(P<0.05),LC3和Beclin1蛋白的表达水平明显下调(P<0.05),而SKOV3细胞无明显变化。结论:卵巢癌DDP耐药性的形成可能与自噬水平增高有关,抑制细胞自噬可以增强SKOV3/DDP细胞对DDP的敏感性。Objective： To investigate the relationship between the autophagy and the sensitivity to DDP （cisplatin） in epithelial ovarian cancer SKOV3 and SKOV3/DDP cells. Methods： The formation of autophagosomes in SKOV3 and SKOV3/DDP cells was observed under a TEM （transmission electron microscope）. MTT assay was used to screen the dose of nontoxic reaction of autophagy inhibitor 3-MA （3-methyladenine） and to examine the sensitivity to DDP in SKOV3 and SKOV3/DDP cells after treatment with the combination of DDP and 3-MA. The apoptotic rate and the expressions of autophagic marker protein LC3 （microtubule-associated protein 1 light chain 3） and autophagy-related protein Beclinl in SKOV3 and SKOV3/DDP cells after treatment with DDP and 3-MA alone and the combination of DDP and 3-MA were detected by flow cytometry and Western blotting, respectively. Results： Much more autophagosomes in SKOV3/DDP cells were observed as compared with that in SKOV3 cells. As compared with DDP treatment alone, when DDP was used in combination with 3-MA, the ICs0 （half inhibitory concentration） value of DDP for SKOV3/DDP cells was decreased from （1 7.71 ±0.1 5） IJg/mL to （8.85± 0.11） IJg/mL （P 〈 0.05）, and the apoptotic rate was increased from （6.93___0.94） % to （14.82±0.85） % （P 〈 0.05）, as well as the expression levels of LC3 and Beclinl proteins in SKOV3/DDP cells were significantly decreased （P 〈 0.05）, but there were no significant changes could be observed in SKOV3 cells. Conclusion： The increased activity of autophagy is related to DDP-resistance of ovarian cancer cells. The inhibition ofautophagy may enhance the sensitivity to DDP in SKOV3/DDP cells.

关 键 词：卵巢肿瘤 自噬 顺铂 耐药 3-MA 

分 类 号：R737.31[医药卫生—肿瘤]