姜黄素对人肾癌Caki-2细胞生长和凋亡的影响及其机制

The Mechanism and Effects of Curcumin on Proliferation Inhibition and Apoptosis Induction in Caki-2 Human Renal Carcinoma Cell

目的:观察姜黄素对体外培养的人肾癌Caki-2细胞的生长抑制作用和凋亡诱导作用,并探讨其可能的作用机制。方法:应用不同浓度的姜黄素分别作用于人肾癌Caki-2细胞12、24、36、48、72、96 h,采用MTT法评价姜黄素对Caki-2细胞的生长抑制作用;应用不同浓度的姜黄素分别作用于人肾癌Caki-2细胞36、48、72、96 h,采用Annexin V-FITC/PI凋亡试剂盒评价姜黄素对Caki-2细胞的凋亡诱导作用;Western blot检测姜黄素对Caki-2细胞H-Ras、p-Raf-B、p-MEK1/2、p-ERK1/2、ERK1/2、Bcl-2、Bax蛋白表达的影响,β-actin作为内参蛋白。结果:不同浓度的姜黄素能显著抑制Caki-2细胞生长,36、48、72、96 h呈剂量、时间依赖关系;不同浓度的姜黄素能显著诱导Caki-2细胞凋亡,48、72、96 h呈剂量、时间依赖关系;姜黄素作用后,Caki-2细胞中H-Ras、p-Raf-B、p-MEK1/2、p-ERK1/2、Bcl-2蛋白表达明显减少,Bax蛋白表达显著增加,ERK1/2、β-actin蛋白表达与空白组相比无统计学差异。结论:姜黄素对体外培养的人肾癌Caki-2细胞有显著的生长抑制作用,其机制可能与通过抑制H-Ras蛋白表达、直接或间接下调ERK信号通路蛋白表达相关;同时,姜黄素对Caki-2细胞有显著的凋亡诱导作用,其机制可能与上调促凋亡蛋白Bax表达、下调抗凋亡蛋白Bcl-2表达相关。

Objective：To study the mechanism and effects of curcumin on proliferation inhibition and apoptosis induction in Caki - 2 human renal carcinoma cell, the experiment was carried out. Methods ： The Caki - 2 human renal carcinoma cell was incu- bated in the presence of different concentrations of curcumin for 12,24,36,48,72 and 96 hours respectively. The inhibitory effect of eurcumin on the proliferation of Caki - 2 human renal carcinoma cell was tested by MTT. The Caki - 2 human renal carcinoma cell was incubated in the presence of different concentrations of curcumin for 36,48,72 and 96 hours respectively. The induction effect of curcumin on the apoptosis of Caki - 2 human renal carcinoma cell was tested by annexin V - FITC/PI apoptosis detection kit. The expressions of H - Ras,p - Raf - B,p - MEK1/2 ,p - ERK1/2, ERK1/2, Bcl - 2 and Bax in Caki - 2 human renal carci- noma cell treated with or without cureumin were tested by western blot. The 13 - actin protein was used to be an internal reference protein. Results ： Curcumin can inhibit Caki - 2 cell proliferation in a dosage and time - dependent manner. Curcumin can induce Caki -2 cell apoptosis in a dosage and time - dependent manner. The expressions of H - Ras, p - Raf - B, p - MEK1/2, p - ERK1/2 and Bcl -2 in Caki -2 cell treated with curcumin were significantly reduced. The expressions of Bax in Caki -2 cell treated with curcumin were significantly increased. There＇s no statistically significant difference between the expressions of ERK1/ 2 and 13 - actin in Caki - 2 cell treated with or without curcumin. Conclusion ： Curcumin can inhibit Caki - 2 cell proliferation in vitro,which may be related to the inhibition of the expressions of H - Ras and down - regulation of ERK signaling pathway in Caki - 2 cell treated by curcumin. Curcumin can induce Caki - 2 cell apoptosis in vitro, which may be related to the up - regulation of expressions of pro - apoptotie protein Bax, and down - regulation of expressions of anti - apoptotic protein Bel - 2 in Caki - 2 cell treated by curcumin.