调节细胞自噬增强卵巢癌顺铂敏感性的体外研究

目的:探讨细胞自噬与上皮性卵巢癌顺铂耐药的关系及调节细胞自噬对卵巢癌顺铂耐药的影响。方法:应用MTT法、MDC荧光染色法及激光共聚焦显微镜技术,观察SKOV3、SKOV3/DDP细胞株自噬现象;通过自噬特异性抑制剂3-MA作用,了解3-MA对卵巢癌细胞胞浆内自噬体的抑制作用及抑制细胞自噬对卵巢癌顺铂敏感性的影响。结果:SKOV3细胞株顺铂IC50值为3.330μg/ml,SKOV3/DDP细胞株顺铂IC50值为14.152μg/ml。3-MA联合顺铂作用后SKOV3细胞株的顺铂IC50值为3.312μg/ml,其对顺铂的敏感性无明显影响(P=0.221),SKOV3/DDP细胞株顺铂IC50值为6.386μg/ml,提示3-MA可提高上皮性卵巢癌顺铂耐药细胞对顺铂的敏感性(P=0.000)。MDC荧光染色及激光共聚焦显微镜观察SKOV3、SKOV3/DDP细胞株胞浆自噬体的形成情况,顺铂能够诱导自噬体的生成,在3-MA作用下,SKOV3/DDP细胞株顺铂诱导的自噬体荧光强度变小,而SKOV3细胞株自噬体的荧光强度变化不大。结论:细胞自噬活性增强与上皮性卵巢癌顺铂耐药有关,抑制细胞自噬活性可以部分逆转上皮性卵巢癌SKOV3/DDP顺铂耐药,但不改变顺铂敏感型细胞SKOV3对顺铂的敏感性。

Adjusting autophagy to augment sensitivity of cisplatin in ovarian carcinoma in vitro

Objective:To explore the relationship between autophagy and cisplatin-resistance in epithelial ovarian cancer and inhibition of autophagy to reverse cisplatin-resistance in epithelial ovarian cancer.Methods:Taifan blue dye,MTT,MDC fluorescence staining and confocal laser scanning microscopy were used to detect the formation of autophagosomes in SKOV3 and SKOV3/DDP.3-MA was used to inhibit activity of autophagy and augment DDP sensitivity in SKOV3/DDP.Results:IC50 of SKOV3 and SKOV3/DDP was 3.330μg/ml and 14.152μg/ml,respectively.When DDP was used combined with 3-MA,IC50 of SKOV3 and SKOV3/DDP was 3.312μg/ml and 6.386μg/ml.There was no influence of 3-MA on DDP sensitivity of SKOV3(P=0.221),but 3-MA could augment DDP sensitivity in SKOV3/DDP(P=0.000).With MDC fluorescence staining and confocal laser scanning microscopy,DDP induced formation of autophagosomes.3-MA decreased the fluorescent density and MDC-labeled particles induced by DDP in SKOV3/DDP,but there was no change in SKOV3.Conclusion:Autophagy is connected with cisplatin-resistance in epithelial ovarian carcinoma.Inhibition of autophagy can partially reverse cisplatin-resistance in SKOV3/DDP,but there is no influence on SKOV3.