siRNA干扰URG11表达对骨肉瘤细胞系MG63的生物学功能和Wnt/β-catenin信号通路的影响

Effects of siRNA mediated interference with URG11 expression on the biological function of osteosarcoma cell line MG63 and Wnt/β-catenin signalling pathway

目的探讨siRNA干扰URG11表达后骨肉瘤细胞系MG63的生物学功能和Wnt/β-catenin信号通路的变化。方法将体外培养的MG63细胞分为Control组(未转染)、NC-siRNA组(转染非特异性NC-siRNA)和URG11-siRNA组(转染URG11-siRNA),采用逆转录聚合酶链式扩增(RT-PCR)检测各组细胞中URG11 mRNA的表达,分别用CCK-8法、Transwell实验和流式细胞仪检测细胞增殖、侵袭和凋亡,用Western blot法检测各组细胞中URG11蛋白以及Wnt/β-catenin信号通路中周期性调控因子D1(Cyclin D1)、原癌基因(c-Myc)、基质金属蛋白酶-2(MMP-2)和凋亡抑制基因(Survivin)的表达。结果与Control组相比,URG11-siRNA转染可使MG63细胞中URG11 mRNA和URG11、β-catenin、c-Myc、Cyclin D1、MMP-2、Survivin蛋白的表达水平降低,同时细胞增殖、侵袭能力显著减弱,细胞凋亡明显增强(均P<0.05);而NC-siRNA转染后MG63细胞的变化无统计学意义(均P>0.05)。结论干扰URG11表达可抑制MG63细胞增殖和侵袭,并促进细胞凋亡,其作用机制可能与抑制Wnt/β-catenin信号通路的活化有关。

Objective To investigate the effects of siRNA mediated interference with URG11 expression on the biological function of osteosarcoma cell line MG63 and Wnt/β-catenin signalling pathway.Methods MG63 cells were divided into control group(un-transfected),NC-siRNA group(transfected with non-specific NC-siRNA)and URG11-siRNA group(transfected with URG11-siRNA).The expression of URG11 was detected by RT-PCR.The proliferation,invasion and apoptosis of MG63 cells were determined by CCK-8 assay,Transwell assay and flow cytometry,respectively.The protein expressions of URG11 and molecules in the Wnt/β-catenin signalling pathway(β-catenin,c-Myc,Cyclin D1,MMP-2 and Survivin)were detected by Western blot method.Results Compared with the control group,URG11-siRNA transfection significantly reduced the mRNA and protein expressions of URG11 as well as the protein expressions of URG11,β-catenin,c-Myc,Cyclin D1,MMP-2 and Survivin in MG63 cells,which was accompanied by the decreased proliferation,inhibited invasion and enhanced apoptosis of MG63 cells(all P<0.05).There were no significant changes in the biological function and Wnt/β-catenin signalling pathway in the MG63 cells after NC-siRNA transfection.Conclusions Interfering the expression of URG11 can inhibit the proliferation and invasion of MG63 cells and promote cell apoptosis,and the mechanism may be related to the inhibition of the activation of Wnt/β-catenin signaling pathway.