【摘要】 目的:研究姜黄素(curcumin)对人肾透明细胞癌(clear-cell renal cell carcinoma, ccRCC)细胞的增殖抑制和诱导凋亡作用，及叉头框蛋白O3a(Forkhead box protein O3a, FOXO3a)在姜黄素抗肿瘤效应中发挥的关键作用。方法:MTT法用于检测姜黄素对ccRCC细胞活力的抑制作用；Annexin V/PI双染实验检测姜黄素对ccRCC细胞的诱导凋亡作用；免疫印迹法(Western blot, WB)检测姜黄素对FOXO3a、p-FOXO3a及下游BIM表达的影响，和凋亡蛋白caspase 3的激活情况。实时荧光定量PCR技术(real-time quantitative PCR,RT-qPCR)检测FOXO3a转录水平。小干扰RNA(small interfering RNA,siRNA)技术用于沉默ccRCC细胞系中FOXO3a的表达。结果:通过对6种ccRCC细胞系(ACHN、786-O、769-P、A-498、Caki-1及Caki-2)FOXO3a蛋白基础表达水平进行检测，选取FOXO3a表达较高的786-O细胞系为研究对象。0～160μmol/L姜黄素处理786-O细胞24 h和48 h后，呈时间和浓度依赖性对细胞活力具有显著抑制作用；786-O细胞在0～80μmol/L姜黄素处理24 h后，明显促进细胞凋亡水平，且伴随着凋亡相关蛋白caspase 3的激活。786-O细胞中，姜黄素抑制p-FOXO3a,促进FOXO3a活化，并上调下游BCL-2家族促凋亡蛋白BIM表达水平。在siRNA沉默FOXO3a蛋白表达后，姜黄素对786-O细胞的生长抑制和诱导凋亡作用被抑制。结论:姜黄素通过诱导激活FOXO3a,在ccRCC细胞上发挥生长抑制和诱导凋亡的抗肿瘤作用。更多还原

【Abstract】 Objective:To evaluate the anti-proliferation and pro-apoptosis effects of curcumin on clear-cell renal cell carcinoma(ccRCC) cells and the role of Forkhead box protein O3(FOXO3 a) played on the antitumor effects of curcumin.Methods:The cell viability inhibition effect of curcumin was assessed by MTT assay.Annexin V/PI staining indicated the curcumin-induced cell apoptosis rates.For mechanism research, FOXO3 a, p-FOXO3 a and FOXO3 a-targeted protein BIM expression,as well as the caspases 3 activation were detected by Western blot. Real-time quantitative PCR( RT-q PCR) was used to determine FOXO3 a transcription. Small interfering RNA( siRNA)was used for FOXO3 a knockout. Results: The basic expression levels of FOXO3 a protein in ccRCC cell lines including ACHN,786-O,769-P,A-498,Caki-1 and Caki-2 were determined by Western blot. 786-O cells with high FOXO3 a expression was chosen for further investigation. 0 ～ 160 μmol/L curcumin inhibited cell viability in a time-and concentration-dependent manner in 786-O cells. The apoptosis rates in cells treated with curcumin increased significantly at 24 h,correlated with the activation of caspases 3. Furthermore,curcumin promoted FOXO3 a activation via decreasing p-FOXO3 a expression,and enhanced the expression of BCL-2 family pro-apoptotic protein BIM,the downstream target protein of FOXO3 a. The proliferation inhibition and apoptosis effects of curcumin in786-O cells could be decreased after FOXO3 a knockout by siRNA. Conclusion: Curcumin played the role of proliferation inhibition and cell apoptosis on 786-O cells via FOXO3 a activation.更多还原