沉默FOSL2基因对骨肉瘤MG63细胞增殖、凋亡、侵袭和迁移的影响

Effects of silencing FOSL2 gene on proliferation,apoptosis,invasion and migration of osteosarcoma cells

目的:探讨沉默FOS样抗原2(FOSL2)基因表达对骨肉瘤细胞生物学特性的影响及机制。方法:采用Western blot检测骨肉瘤和肉瘤旁正常组织中FOSL2蛋白的表达。取对数生长期MG63细胞分为空白对照组(加生理盐水)、阴性对照组(NC组,转染阴性对照siRNA)和FOSL2-siRNA组(转染FOSL2 siRNA),通过CCK-8、Annexin V-FITC/PI双染法及Transwell小室分别检测各组细胞增殖、凋亡情况和侵袭、迁移能力,Western blot检测FOSL2、PCNA、MMP-2和Bax蛋白表达,qRT-PCR检测PCNA、MMP-2和Bax mRNA表达。结果:FOSL2-siRNA组MG63细胞中FOSL2蛋白表达明显低于空白对照组和NC组(P<0.05)。与空白对照组和NC组比较,FOSL2-siRNA组细胞增殖明显降低,凋亡率明显升高,细胞侵袭和迁移能力明显降低,MMP-2和PCNA表达明显降低,Bax表达明显升高(P<0.05)。结论:沉默FOSL2基因表达可抑制骨肉瘤细胞增殖、侵袭和迁移能力,并诱导细胞凋亡,机制与下调PCNA和MMP-2表达,上调Bax表达有关。

Aim:To investigate the effects of silencing FOSL2 gene on proliferation,apoptosis,invasion and migration of osteosarcoma cells and its mechanism.Methods:Western blot was used to detect FOSL2 expression in the osteosarcoma and normal tissue.MG63 cells in the logarithmic growth phase were divided into blank group(adding normal saline),negative control group(NC group,transfecting negative control siRNA)and FOSL2-siRNA group(transfecting FOSL2 siRNA).CCK-8 assay,Annexin V-FITC/PI double staining and Transwell chamber were used to detect cell proliferation,apoptosis,invasion and migration ability.Western blot was used to detect the expressions of FOSL2,PCNA,MMP-2 and Bax proteins.The expressions of PCNA,MMP-2 and Bax mRNA were detected by qRT-PCR.Results:The expression of FOSL2 protein in the FOSL2-siRNA group was significantly lower than those in the blank group and the NC group(P<0.05).Compared with the blank group and the NC group,cell proliferation ability in the FOSL2-siRNA group decreased,apoptosis rate increased,cell invasion and migration ability decreased,MMP-2 and PCNA expression decreased,and Bax expression increased(P<0.05).Conclusion:Silencing FOSL2 gene expression can inhibit proliferation,invasion and migration of osteosarcoma cells,and induce apoptosis,which is related to down-regulation of PCNA and MMP-2 expression and up-regulation of Bax expression.