沉默RIPK4基因表达可抑制骨肉瘤U-2OS细胞上皮-间质转化的发生

RIPK4 gene-silencing inhibits epithelial-mesenchymal transition of osteosarcoma U-2OS cells

目的 :探讨沉默受体相互作用蛋白激酶4(receptor interacting protein kinase4,RIPK4)基因表达对骨肉瘤U-2OS细胞上皮-间质转化(epithelialmesenchymal transition,EMT)的影响。方法 :采用脂质体法将特异性针对RIPK4基因的si RNA转入骨肉瘤U-2OS细胞,构建RIPK4基因沉默表达的U-2OS细胞系,随后采用蛋白质印迹法检测RIPK4表达的变化。采用Transwell小室法检测沉默RIPK4基因表达后对U-2OS细胞迁移和侵袭能力的影响;倒置光学显微镜下观察细胞形态的变化。最后,釆用蛋白质印迹法检测U-2OS细胞中EMT标志物E-钙黏蛋白(E-cadherin)和波形蛋白(vimentin)的表达水平。结果:蛋白质印迹法检测结果显示,RIPK4-si RNA转入U-2OS细胞后,RIPK4蛋白的表达水平明显下调(P<0.05);RIPK4-si RNA转染组U-2OS细胞的迁移和侵袭能力明显下降(P值均<0.05);U-2OS细胞的形态由间质形向上皮细胞形转变。RIPK4-si RNA转染组细胞中E-cadherin的表达水平明显上调(P<0.05),而vimentin的表达水平明显下调(P<0.05)。结论 :沉默RIPK4基因表达可抑制骨肉瘤U-2OS细胞EMT的发生,并降低细胞的迁移和侵袭能力。

Objective： To investigate the effects of receptor interacting protein kinase 4（RIPK 4） gene-silencing on epithelial-mesenchymal transition（EMT） of osteosarcoma U-2OS cells.Methods： The specific si RNA targeting RIPK 4 gene was transfected into U-2OS cells by liposome to establish RIPK 4 gene-silencing U-2OScell line. Then the change of RIPK4 expression level was detected by Western blotting. The migration and invasion abilities of U-2OS cells after RIPK 4 gene-silencing were detected by Transwell chamber assay. The morphological changes of U-2OS cells were observed under an inverted optical microscope. Finally, the expression levels of EMT makers E-cadherin and vimentin in U-2OS cells were detected by Western blotting.Results： After transfection with RIPK4-si RNA, the expression level of RIPK4 protein was significantly decreased in U-2OS cells（P〈0.05）. The migration and invasion abilities of U-2OS cells in RIPK4-si RNA transfection group were significantly reduced（both P〈0.05）. The morphology of U-2OS cells conversed from mesenchymal phenotype to epithelial phenotype. The expression level of E-cadherin was significantly up-regulated in RIPK4-si RNA transfection group（P〈0.05）, whereas the expression level of vimentin was significantly down-regulated（P〈0.05）.Conclusion： RIPK 4 gene-silencing can inhibit the occurrence of EMT in osteosarcoma U-2OS cells, and reduce the migration and invasion abilities of osteosarcoma cells.