姜黄素抑制人肾癌细胞769-P增殖及促凋亡的实验研究

Effect of curcumin on the proliferation and apoptosis of human renal cell of carcinoma cell line769-P

目的 分析不同浓度姜黄素（curcumin,分子式C21H20O6）对人肾癌细胞769-P的生物学作用及其调控机制.方法 体外培养人肾癌细胞769-P,用不同浓度姜黄素处理24 h后,采用MTT比色法测定姜黄素对细胞的增殖抑制作用,利用倒置显微镜观察细胞的显微形态结构,采用细胞划痕法观察细胞的迁移情况;Western Blot法检测细胞内Caspase-3、AIF蛋白质的表达情况;用反转录PCR法检测AIF、Bax mRNA的表达水平.结果 MTT法检测显示姜黄素能显著抑制人肾癌细胞系769-P的增殖,呈现浓度和时间效应关系;倒置显微镜结果显示姜黄素能够使肾癌细胞系的形态学发生改变,呈现出典型凋亡特征如核皱缩、核破裂、核肿胀等形态;细胞划痕实验显示,姜黄素抑制了肾癌细胞系的迁移;RT-PCR实验显示,姜黄素促进了BAX mRNA和AIF mRNA的表达;蛋白杂交实验显示,姜黄素促进了Caspase-3和AIF蛋白的活化,从而促进了凋亡.结论 姜黄素能显著抑制人肾癌细胞的增殖并促进其凋亡,这种生物学效应可能与激活Bax和AIF表达,活化Caspase-3的信号通路有关.

Objective To explore the biological function of different concentrations of curcumin on human renal carcinoma cell line 769-P,and to investigate its mechanisms.Methods Human renal carcinoma cell line 769-P in vitro was cultured,then different concentration of curcumin for 24 hours was treated respectively.The MTT assay was used to measure the inhibition effect of curcumin on cell proliferation.The inverted microscope was utilized to observe the microscopic morphology of cell.Cell scratch method was used to detect the migration of cell.The expression of Caspase-3 and AIF protein were evaluated by Western Blot.The expressions of AIF and Bax mRNA were evaluated by reverse transcription polymerase chain reaction.Results MTT results showed that curcumin could significantly inhibit the proliferation of human renal carcinoma cell line 769-P,with dose-and time-dependent effects.Inverted microscope results showed that curcumin could change the morphological feature of renal carcinoma cell line,with typical apoptotic morphological feature such as nuclear shrinkage,nuclear burst,nuclear swelling and so on.Cell scratch results showed that curcumin inhibit the renal carcinoma cell migration.RT-PCR results showed that curcumin up-regulated the expression of BAX and AIF mRNA level.Western Blot results showed that curcumin promoted the activation of caspase-3 and AIF protein,so as to promote the apoptosis.Conclusions Curcumin can significantly inhibit the proliferation of human renal carcinoma cell and promoted its apoptosis.This biological effect may be related to the activation of Bax and AIF and activation of caspase-3 signaling pathways.