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目的研究化学合成的靶向MIF的siRNA干扰MIF后,对大肠癌CT-26细胞增殖的影响并探讨其可能的机制。方法 MTT法检测细胞的增殖情况;ELISA检测培养上清液中MIF蛋白的含量;逆转录聚合酶链反应(RT-PCR)检测MIF、CD74 mRNA的表达;Western blot检测细胞内MIF和CD74蛋白的表达。结果实验组CT-26细胞的增殖与对照组和空白组相比受到明显抑制(P24 h;100 nmol=0.003),呈时间-计量依赖关系;实验组培养上清液中MIF蛋白的含量与对照组和空白组比较显著减少(P=0.02);实验组中MIF与CD74的mRNA表达量与对照组和空白组相比显著下降(PMIF=0.001;PCD74=0.001);实验组MIF、CD74蛋白表达与对照组和空白组相比降低(PMIF=0.006;PCD74=0.016)。结论化学合成的MIF siRNA抑制了CT-26细胞的增殖,其可能机制是MIF siRNA降低了CT-26细胞内MIF与CD74的表达。
Objective To investigate the effect of chemically synthesized siRNA targeting MIF on the proliferation of colorectal cancer CT-26 cells and to explore its possible mechanism. Methods MTT method was used to detect the proliferation of MIF and CD74 mRNA in culture supernatant. The expression of MIF and CD74 mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) expression. Results The proliferation of CT-26 cells in the experimental group was significantly inhibited compared with the control group and the blank group (P24 h; 100 nmol = 0.003) in a dose-dependent manner. The content of MIF protein in the culture supernatant of the experimental group was significantly lower than that of the control group (P = 0.02). The mRNA expression of MIF and CD74 in the experimental group was significantly decreased compared with the control group and the blank group (PMIF = 0.001; PCD74 = 0.001). The expression of MIF and CD74 in the experimental group Decreased compared with the control group and the blank group (PMIF = 0.006; PCD74 = 0.016). Conclusion Chemically synthesized MIF siRNA inhibits the proliferation of CT-26 cells. The possible mechanism is that MIF siRNA reduces the expression of MIF and CD74 in CT-26 cells.