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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 investigate the apoptosis of human renal cell carcinoma 786-O cell induced by tanshinone ⅡA and its molecular mechanism. Methods: The effect of Tanshinone ⅡA on the viability of 786-O cells was detected by MTT assay. Apoptosis induction of 786-O cells by tanshinone ⅡA was detected by flow cytometry. The expression of target protein was detected by Western blotting. Results: MTT assay showed that the survival rates of 786-O cells were 100.0%, 68.4%, 46.3%, 33.5% and 28.2% after treatment with tanshinone Ⅱ A at concentrations of 0, 1, 2, 4 and 8 μg / The proliferation of 786-O cells was significantly inhibited in a concentration-dependent manner (P <0.05). AnnexinⅤ / PI double staining and flow cytometry showed that after treatment with 0,2,4 and 8μg / mL tanshinoneⅡA for 24 hours, O cells were 12.3%, 36.4%, 42.1% and 43.9% respectively, indicating that tanshinone IIA can induce 786-O cell apoptosis in a concentration-dependent manner. Western blotting showed that tanshinone IIA-treated 786-O cells , p53 and its downstream target gene Bax were significantly upregulated, and activated Caspase-3. Conclusion: Tanshinone Ⅱ A can induce apoptosis of 786-O cells by up-regulating p53 and its downstream gene Bax, and then activating Caspase-3.