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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。
Objective: To investigate the relationship between autophagy and cisplatin resistance in epithelial ovarian cancer and the regulation of autophagy on cisplatin resistance in ovarian cancer. Methods: The autophagy of SKOV3 and SKOV3 / DDP cell lines was observed by MTT assay, MDC fluorescence staining and laser confocal microscopy. The effect of 3-MA on ovarian cancer cell Effect of inhibition of autophagosome in plasma and inhibition of cell autophagy on cisplatin sensitivity in ovarian cancer. Results: The IC50 of cisplatin in SKOV3 cell line was 3.330μg / ml, and the IC50 of cisplatin in SKOV3 / DDP cell line was 14.152μg / ml. The IC50 value of cisplatin of SKOV3 cell line after 3-MA combined with cisplatin was 3.312μg / ml, which had no significant effect on the sensitivity of cisplatin (P = 0.221). The IC50 of cisplatin of SKOV3 / DDP cell line was 6.386μg / ml, suggesting that 3-MA can improve cisplatin sensitivity of epithelial ovarian cancer cisplatin-resistant cells (P = 0.000). The formation of autophagosomes in SKOV3 and SKOV3 / DDP cell lines was observed by MDC fluorescent staining and confocal laser scanning microscope. Cisplatin could induce the formation of autophagosomes. Under the action of 3-MA, cisplatin was induced in SKOV3 / DDP cell lines The fluorescence intensity of autophagosome in SKOV3 cell line did not change much. CONCLUSION: The enhanced autophagy activity is associated with cisplatin resistance in epithelial ovarian cancer. Inhibition of autophagic activity may partially reverse cisplatin-resistant SKOV3 / DDP cells, but not cisplatin-sensitive cells Sensitivity.