单宁酸对肾癌细胞生长的抑制作用及其机制

Inhibitory effect of tannic acid on growth of renal cell carcinoma cells and its mechanism

目的:观察单宁酸对人肾癌细胞OS-RC-2增殖及凋亡的影响,并初步探讨其作用机制。方法:培养人肾癌细胞OS-RC-2,将其分为空白对照组、不同浓度TA组(20、40、80和100μmol/L),观察OS-RC-2的细胞形态,CCK8法检测细胞增殖,流式细胞术检测细胞凋亡,DCF法测定细胞内ROS,化学显示法检测抗氧化酶活性,ELISA法细胞上清中ICAM-1蛋白表达,Westernblot法检测Bcl-2、Bax及Caspase-3蛋白的表达。结果:TA可有效抑制OS-RC-2肾癌细胞增殖,并促进其凋亡,呈显著剂量效应关系。TA可诱导OS-RC-2肾癌细胞内ROS生成增加,增加细胞中MDA含量,降低GSH-Px及SOD的活性,降低OS-RC-2细胞ICAM-1蛋白的分泌,下调细胞核中Bcl-2蛋白表达及上调Bax及Caspase-3凋亡蛋白的表达。结论:TA可能通过干预肾癌细胞内ROS及氧化酶的活性、抑制ICAM-1蛋白分泌及靶向线粒体凋亡等机制参与抑制肾癌细胞生长及促进其凋亡。

Objective: To observe the effect of tannic acid (TA) on the proliferation and apoptosis of human renal cell carcinoma OS-RC-2,and to explore its mechanism. Methods: Human renal cancer cell OS-RC-2 was cultured and divided into blank control group and TA group (20,40,80 and 100 μmol/L).The cell morphology of OS-RC-2 was observed.Cell proliferation was detected by CCK8 method.Cell apoptosis was detected by flow cytometry.ROS was detected by DCF method.Antioxidant enzyme activity was detected by chemical display method.ICAM-1 protein expression in cell supernatant was detected by ELISA method.The expression of Bcl-2,Bax and Caspase-3 protein was detected by Western blot method. Results: TA can effectively inhibit the proliferation and promote the apoptosis of OS-RC-2 renal cancer cells in a dose-dependent manner.TA can induce ROS production in OS-RC-2 renal cell carcinoma cells,increase the content of MDA,decrease the activity of GSH-Px and SOD,decrease the secretion of ICAM-1 protein in OS-RC-2 cells,down-regulate the expression of Bcl-2 protein in the nucleus and up-regulate the expression of Bax and Caspase-3 apoptotic protein. Conclusion: TA may be involved in inhibiting the growth and promoting apoptosis of renal cell carcinoma cells by interfering with the activity of ROS and oxidase,while inhibiting the secretion of ICAM-1 protein and targeting mitochondrial apoptosis.