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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 investigate the mechanism of Interferon-γ (IFN-γ) on the proliferation of renal cell carcinoma. Methods Human renal carcinoma 786-0 cells were treated with IFN-γ at the concentration of 1 000, 2 000 and 3 000 U / mL. After 24, 48 and 72 h of treatment, the cell proliferation inhibition rate was determined by CCK-8 assay. Cytokines were analyzed by flow cytometry. The mRNA expression of hepaCAM was detected by RT-PCR. The expression of MAD1 protein was detected by Western blot. Results Different concentrations of IFN-γ had inhibitory effects on the proliferation of renal cancer cells. The inhibitory rates of different concentrations of IFN-γ at different times were 72 h> 48 h> 24 h, with statistical significance (P <0.05) (P <0.05). The results of cell cycle distribution showed that the proliferation of G0 / G1 phase cells in renal cell carcinoma of experimental group was abnormal after 48 h of treatment. Compared with the control group (39.89) Compared with the control group, the proliferation index (25.65) in the experimental group decreased significantly (P <0.05). The results showed that the expression of hepaCAM and MAD1 in the experimental group was significantly increased High, the difference was statistically significant (P <0.05). Conclusion Interferon-γ can improve the expression of hepaCAM, increase the expression of MAD1 and hinder the proliferation of renal cell carcinoma, which is of clinical significance.