过表达miR-205对顺铂诱导的肾上腺皮质癌细胞株SW-13凋亡的影响及其机制

目的 研究过表达miR-205对顺铂（cisplantin，CDDP）诱导的肾上腺皮质癌细胞株SW-13凋 亡的影响及其机制。方法 设转染pcDNA3.1( + )-miR-205的SW-13细胞和转染 pcDNA3.1( + )的SW-13 细胞分别为实验组和对照组。在不同浓度顺铂作用下，采用CCK-8法比较两组细胞对CDDP敏感度； H33342及AnnexinV-FITC/PI荧光观察细胞凋亡，AnnexinV-PE/7AAD荧光流式细胞术检测细胞凋亡 率；用Western blot检测目的蛋白的表达水平；qPCR检测相关基因mRNA水平的变化。结果 不同浓度 顺铂作用下，实验组细胞生长抑制率、凋亡率明显高于对照组（P＜0.01，P＜0.05）；在0、20、30 μg/ml浓度的CDDP处理后，实验组促凋亡蛋白Bax、Caspase-9以及E2F1和VEGF-A mRNA水平较对照 组升高，而抗凋亡蛋白Bcl-2表达则降低。结论 上调SW-13细胞miR-205可降低E2F1、VEGF-A的表 达水平，促进顺铂诱导细胞的凋亡，增强肾上腺皮质癌细胞对顺铂的敏感度。

Effects of miR-205 Overexpression on Cisplatin-induced Apoptosis of Adrenocortical Carcinoma Cell Line SW-13 and Its Mechanism

Objective To investigate the effect of miR-205 overexpression on apoptosis of adrenocortical carcinoma cell line SW-13 induced by cisplatin (CDDP) and to explore the related mechanism. Methods Two kinds of adrenocortical carcinoma cell lines SW-13, one transfected with pcDNA3.1( + )-miR-205(treatment group) and the other transfected with pcDNA3.1( + ) (control group), were involved. With different concentrations of cisplantin, sensitivities of two groups to different doses of CDDP were tested by CCK-8 assay, apoptosis was examined by the staining of H33342 and AnnexinV-FITC/PI, AnnexinVPE/ 7AAD flow cytometry was applied to assess apoptosis rates, expression levels of apoptosis-related proteins were detected by Western blot, and mRNA expression levels of E2F1 and VEGF-A were investigated by qPCR. Results The apoptosis and growth inhibition rates in treatment group were significantly higher than those in control group（P＜0.05, P＜0.01）. After treated with 0, 20, 30μg/ml of CDDP, Bcl-2 expression and mRNA expression of E2F1 and VEGF-A in treatment group were decreased, while Bax, Caspase-9 expression in treatment group were markedly increased. Conclusion MiR-205 overexpression in SW-13 would down-regulate E2F1 and VEGF-A expression levels, enhance the sensitivity of adrenocortical carcinoma cells to cisplantin, and promote cell apoptosis.