siRNA下调THBS4对人骨肉瘤MG63增殖的影响

The effect of knock-down of THBS4 by siRNA on the proliferation of human osteosarcoma cell line MG63

目的探讨小干扰RNA(small interfering RNA,siRNA)抑制血小板反应蛋白4(thrombospondin 4,THBS4)基因的表达对骨肉瘤MG-63细胞增殖的影响及其分子机制。方法将合成的THBS4 siRNA转染到人成骨细胞株MG63,CCK-8法观察对细胞增殖的影响,荧光定量PCR检测THBS4及TGF-beta1的mRNA表达,Western-blotting检测THBS4及TGF-beta1的蛋白表达。结果 Thbs4 siRNA转染后实验组48 h,72 h OD值高于阴性对照组(P<0.05),差异具有显著性;Thbs4 mRNA和蛋白的表达水平明显低于阴性对照组和空白对照组(P<0.01);Thbs4 siRNA转染后MG63细胞中TGF-beta1 mRNA及蛋白的表达水平提高(P<0.01)。结论 THBS4 siRNA转染MG63后能够促进细胞增殖,其机制可能是激活TGF-beta1信号通路。

Objective To investigate the effect of thrombospondin 4(THBS4)gene silencing by small interfering RNA(siRNA)on the proliferation of MG63 and to explore its molecular mechanism.Methods After THBS4 siRNA was transfected into MG63 cells,the proliferation was detected with CCK-8 method.The mRNA expression THBS4 and TGF-beta1 was measured using real-time fluoresce quantitative-PCR.The protein levels of THBS4 and TGF-beta1 were detected with Western blotting.Results The optical density in experimental group at 48 h and 72 h was significantly higher than that in the negative control group(P<0.05).The expression levels of THBS4 mRNA and protein in MG63 cells after transfection with THBS4 siRNA were lower than those in the negative control cells and the blank control cells(P<0.01).The expression levels of TGF-beta1 mRNA and protein in MG63 cells after transfection were up regulated.Conclusion THBS4 siRNA transfection promotes MG63 proliferation through the activation of TGF-beta1 signaling pathway.