丹参酮ⅡA诱导肾癌786-O细胞凋亡及其分子机制的研究

Apoptotic effect and its molecular mechanism of Tanshinone ⅡA on human renal carcinoma 786-O cells

目的:研究丹参酮ⅡA诱导肾细胞癌786-O细胞凋亡及其分子机制。方法:MTT法检测丹参酮ⅡA对786-O细胞活力影响;流式细胞分析检测丹参酮ⅡA对786-O细胞凋亡诱导;蛋白质印迹法检测诱导凋亡相关靶蛋白表达水平。结果:MTT分析显示,用浓度0、1、2、4及8μg/mL丹参酮ⅡA处理24h后,786-O细胞生存率分别为100.0%、68.4%、46.3%、33.5%和28.2%,表明丹参酮能够呈浓度依赖方式显著抑制786-O细胞生长活力,P<0.05;AnnexinⅤ/PI双染法,流式细胞仪检测显示,采用浓度0、2、4及8μg/mL丹参酮ⅡA处理24h后,786-O细胞凋亡率分别为12.3%、36.4%、42.1%和43.9%,表明丹参酮ⅡA能够以浓度依赖方式诱导786-O细胞凋亡;蛋白质印迹法检测结果表明,丹参酮ⅡA处理786-O细胞,p53及其下游靶基因Bax显著上调,并激活Caspase-3。结论:丹参酮ⅡA诱导786-O细胞凋亡,其机制可能通过上调p53及其下游基因Bax,继而激活Caspase-3。

OBJECTIVE： To explore the apoptotic effect and its molecular mechanism of Tanshinone μ A on human renal cell carcinoma 786-0 cells. METHODS： MTT assay was used to detect vitality of 786-0 cells treated with Tanshi- none ⅡA;Flow Cytometry was exployed to examine apoptosis of 786-0 cells induced by TanshinoneⅡ A;Immunoblotting was utilized to determine the expression levels of target proteins related to apoptosis. RESULTS.. MTT assay showed the percentages of 786-0 viable cells were 100.0% ,68.4%,46.3% ,33.5% and 28.2% when cultured with TanshinoneⅡA （0,1,2,4 and 8 μg/mL,respectively） for 24 h. The result indicated Tanshinone ⅡA remarkably impaired 786-0 cells vitality in dose-dependent manner （P〈0. 05）. Annexin V and propidium iodide staining showed that the apoptotic rates of 786-0 cells following the treatment of 786-0 cells for 24 h with Tanshinone ⅡA （0,2,4 and 8 μg/mL, respectively） were 12.3%,36. 4%, 42. 1% and 43. 9%, indicating Tanshinone ⅡA induced significant apoptosis in 786-0 cells. Further more,Immunoblotting indicated the upregulation of p53 plus its target gene bax,and the activation of caspase-3 in 786-0 cells treated with Tanshinone H A. CONCLUSION： Tanshinone ⅡA induced apoptosis 786-0 cells maybe via upregulation of p53 and its target gene bax which caused ensuing activation of caspase-3.