敲低性别决定区Y-box蛋白5对胃癌细胞增殖、侵袭和上皮间质转化的影响

Effects of Knockdown on Sex Determining Region Y-box Protein 5 Gastric Cancer Cell Proliferation, Invasion and Epithelial-Mesenchymal Transition

[目的]探讨敲低性别决定区Y-box蛋白5(Sox-5)的表达水平及对胃癌细胞增殖、侵袭和上皮间质转化(EMT)的影响。[方法]实时荧光定量聚合酶链反应(qRT-PCR)检测胃癌细胞和正常人胃黏膜上皮细胞中Sox-5 m RNA的相对表达水平;sh-NC和sh-Sox-5质粒转染BGC823细胞,蛋白免疫印迹实验检测Sox-5蛋白表达水平;MTT实验、划痕实验和Transwell实验分别检测BGC823细胞增殖活性、迁移能力和侵袭能力;蛋白免疫印迹实验检测扭曲蛋白(Twist)、波形蛋白(Vimentin)、E-钙黏蛋白(E-cadherin)的表达。[结果]胃癌细胞中Sox-5 mRNA的相对表达水平高于正常人胃黏膜上皮细胞(P<0.01);与转染sh-NC组相比,转染sh-Sox5质粒组中Sox-5蛋白的相对表达水平降低(1.120±0.142 vs 0.124±0.010,t=9.032,P<0.001);与转染sh-NC组相比,转染sh-Sox-5组细胞增殖活性显著降低(1.32±0.10 vs 0.74±0.06,t=3.514,P=0.005),迁移率能力减弱(46.2%±4.7%vs 17.6%±2.5%,t=9.625,P<0.001),侵袭能力减弱(93.7±9.1 vs 113.7±6.5,t=3.103,P=0.036);敲低Sox-5蛋白表达后Twist和Vimentin蛋白表达减弱,E-cadherin蛋白表达增强(P均<0.001)。[结论]敲低胃癌细胞中Sox-5的表达,可抑制胃癌细胞的增殖、迁移、侵袭和EMT。

[Objective] To investigate the effect of knocking down expression level of sex determining region Y-box protein 5(Sox-5) on the proliferation, invasion and epithelial-mesenchymal transition(EMT) of gastric cancer cells. [Methods] The relative expression level of Sox-5 m RNA in gastric cancer cells and normal human gastric mucosal epithelial cells was detected by real-time fluorescent quantitative polymerase chain reaction(q RT-PCR). The sh-NC and sh-Sox-5 plasmids were transfected into gastric cancer BGC823 cells, and the protein expression level of Sox-5 was detected by Western blotting. The proliferation activity, migration ability and invasion ability of BGC823 cells was detected by MTT method, scratch assay and transwell test, respectively. The expression of Twist, Vimentin,and E-cadherin proteins was detected by Western blotting. [Results] The relative expression level of Sox-5 m RNA in gastric cancer cells was higher than that in normal human gastric mucosal epithelial cells(P<0.01). Compared with the sh-NC transfected group, the relative expression level of Sox-5 protein in the sh-Sox5 plasmid transfected group was reduced(1.120±0.142 vs 0.124±0.010, t=9.032, P<0.001). Compared with the transfected sh-NC group, the cell proliferation activity of the transfected shSox-5 group was significantly reduced(1.32±0.10 vs 0.74±0.06, t=3.514, P=0.005), the cell migration rate was decreased(46.2%±4.7% vs 17.6%±2.5%, t=9.625, P<0.001), and the invasion ability was also decreased(93.7±9.1 vs 113.7±6.5, t=3.103, P=0.036). After knocking down Sox-5 protein expression, the expression of Twist and Vimentin protein decreased, and the expression of E-cadherin protein increased(all P<0.001). [Conclusion] Knockdown of Sox-5 expression can inhibit the proliferation,migration, invasion and EMT of gastric cancer cells.