干扰素-γ对肾癌细胞增殖干预的机制研究

Study on the mechanism of interferon-γ on the proliferation of renal carcinoma cell intervention

目的研究干扰素-γ(IFN-γ)对肾癌细胞增殖干预的机制。方法应用浓度1 000、2 000、3 000 U/mL的IFN-γ处理肾癌786-0细胞株,在处理24、48、72 h后,采用CCK-8法测定细胞增殖抑制率,采用流式细胞仪分析细胞周期,采用RT-PCR法检测肝细胞粘附分子(hepaCAM)mRNA的表达,采用Western bolt方法检测MAD1蛋白表达情况。结果不同浓度的IFN-γ对肾癌细胞的增殖均有抑制作用,相同浓度不同时间抑制率72h>48 h>24 h,差异有统计学意义(P<0.05);相同作用时间,IFN-γ浓度越大,抑制率越大,差异有统计学意义(P<0.05);细胞周期分布结果显示,实验组肾癌细胞在处理48 h后出现G0/G1期增殖异常;与对照组(39.89)比较,实验组增殖指数(25.65)明显下降,差异有统计学意义(P<0.05);结果显示,实验组肾癌细胞在处理48 h后,与对照组比较,hepaCAM、MAD1蛋白表达量明显升高,差异有统计学意义(P<0.05)。结论干扰素-γ可以通过促进hepaCAM基因表达,使MAD1蛋白表达升高,阻碍肾癌细胞增殖过程,对临床具有指导意义。

Objective To study the mechanism of interferon-γ(IFN-γ)on the intervention of renal carcinoma cell proliferation.Methods Using concentration of 1 000,2 000,3 000 U/mL IFN-γtreatment of renal cell carcinoma 786-0 cell line,in 24 hours,48 hours,72 hours after treatment,the inhibition rate of cell proliferation was determined with CCK-8 method,using flow cytometric analysis of cell cycle,using RT-PCR for detection of hepaCAM mRNA,and using the Western boltting method for detection of MAD1 protein expression.Results Different concentrations of IFN-γhad the inhibitory effects on renal cell carcinoma cell proliferation,the concentration of the inhibitory rate of 72 hoursand 48 hours more than 24 hours,the difference was statistically significant (P<0.05);at the same time,a higher IFN-γconcentration,the inhibition rate was greater,the difference was statistically significant (P<0.05 );the cell cycle distribution results showed,the experimental group of renal carcinoma cells proliferation in the treatment of abnormal G0/G1 phase after 48 hours;and the control group (39.89 )compared with the experimental group,the proliferation index (25.65 )decreased significantly,the difference was statistically significant (P<0.05 );results showed that,the experimental group in renal cell carcinoma cells after 48 h of treatment,compared with control group,hepaCNM mRNA,MAD1 protein expression increased obviously,the difference was statistically significant (P<0.05 ).Conclusion IFN-γcould increase the expression of MAD1 by promoting hepaCAM expression,inhibits renal carcinoma cell proliferation.