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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

聂奇伟, 胡卫列, 欧阳可育, 曹 曙, 王 尉

聂奇伟, 胡卫列, 欧阳可育, 曹 曙, 王 尉. 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[J]. 肿瘤防治研究, 2014, 41(07): 719-723. DOI: 10.3971/j.issn.1000-8578.2014.07.007
引用本文: 聂奇伟, 胡卫列, 欧阳可育, 曹 曙, 王 尉. 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[J]. 肿瘤防治研究, 2014, 41(07): 719-723. DOI: 10.3971/j.issn.1000-8578.2014.07.007
NIE Qiwei, HU Weilie, OUYANG Keyu, CAO Shu, WANG Wei. Effects of miR-205 Overexpression on Cisplatin-induced Apoptosis of Adrenocortical Carcinoma Cell Line SW-13 and Its Mechanism[J]. Cancer Research on Prevention and Treatment, 2014, 41(07): 719-723. DOI: 10.3971/j.issn.1000-8578.2014.07.007
Citation: NIE Qiwei, HU Weilie, OUYANG Keyu, CAO Shu, WANG Wei. Effects of miR-205 Overexpression on Cisplatin-induced Apoptosis of Adrenocortical Carcinoma Cell Line SW-13 and Its Mechanism[J]. Cancer Research on Prevention and Treatment, 2014, 41(07): 719-723. DOI: 10.3971/j.issn.1000-8578.2014.07.007

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

基金项目: 国家自然科学基金资助项目(81172421);广东省自然科学基金资助项目(S2012010010009)
详细信息
    作者简介:

    聂奇伟(1986-),男,硕士在读,主要从事肾上腺肿瘤的研究

    通讯作者:

    胡卫列,E-mail:huwl-mr@vipsina.com

  • 中图分类号: R736.6

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

  • 摘要: 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。

     

    Abstract: 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.

     

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出版历程
  • 刊出日期:  2014-07-24

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