SFRP4表达对胃癌细胞SGC-7901迁移和侵袭的影响及机制

Effect of SFRP4 expression on migration and invasion of gastric cancer cell line SGC-7901 and its mechanism

目的 分析分泌型卷曲相关蛋白4(SFRP4)在胃癌细胞中的表达,探讨SFRP4对胃癌细胞迁移以及侵袭的影响机制。方法 选取正常胃黏膜上皮细胞GES-1和胃癌细胞SGC-7901分别培养,蛋白质印迹法检测SFRP4表达。构建SFRP4对照质粒及敲低质粒转染至SGC-7901细胞,分别作为Scr组和SiSFRP4组,另外设置空白组(SGC-7901),通过蛋白质印迹法检测质粒转染效率;划痕实验以及Transwell侵袭实验分别检测转染质粒后胃癌细胞的迁移以及侵袭能力;蛋白质印迹法检测转染质粒后胃癌细胞中基态金属蛋白酶(MMPs)和上皮间充质转化(EMT)标志物的表达变化。结果 胃癌细胞株SGC-7901 SFRP4蛋白表达水平为2.54±0.19,高于GES-1细胞(1.00±0.11),差异有统计学意义,t=12.150,P<0.001。质粒转染后,Scr组SFRP4表达水平为0.95±0.10,SiSFRP4组为0.27±0.06,差异有统计学意义,t=10.100,P<0.001。SiSFRP4组穿过Transwell小室Matrigel滤膜的细胞数量(0.41±0.10)低于Scr组(1.00±0.12),t=6.542,P=0.003。转染24和48 h后,SiSFRP4组细胞迁移率分别为0.42±0.13和0.41±0.08,低于Scr组的1.00±0.17和1.00±0.08,差异有统计学意义,t值分别为4.693和8.851,P值分别为0.009和<0.001。转染24 h后,SiSFRP4组E-cadherin表达(2.35±0.15)高于Scr组(1.00±0.22),差异有统计学意义,t=8.782,P<0.001;SiSFRP4组Vimentin表达(0.31±0.14)低于Scr组(1.00±0.27),差异有统计学意义,t=3.930,P=0.017;SiSFRP4组MMP-2及MMP-9表达水平分别为0.58±0.09和0.66±0.08,低于Scr组的1.00±0.12和1.00±0.10,差异有统计学意义,t值分别为4.850和4.599,P值分别为0.008和0.010。结论 SFRP4在胃癌细胞中高表达,且可通过EMT途径及上调MMPs表达促进胃癌细胞的迁移以及侵袭过程。

Objective To investigate the expression of secreted frizzled related protein 4(SFRP4) in gastric cancer cells, and explore the influence mechanism of SFRP4 on the migration and invasion process of gastric cancer cells.Methods Normal gastric mucosal epithelial cells GES-1 and gastric cancer cells SGC-7901 were cultured respectively, and the expression of SFRP4 was detected by Western blot assay.SFRP4 control plasmid and knockdown plasmid were constructed and transfected into SGC-7901 cells, which were used as control group(Scr) and interference group(SiSFRP4),respectively.In addition, blank group(SGC-7901) was set, the plasmid transfection efficiency was detected by Western blot.The assays of wound healing and Transwell invasion assay were used to detect the migration and invasion of gastric cancer cells after transfection plasmid.The expression changes of matrix metalloproteinase(MMPs) and epithelial-mesenchymal transition(EMT) markers in gastric cancer cells after transfection plasmid were detected by Western blot.Results The expression level of SFRP4 protein in gastric cancer cell line SGC-7901(2.54±0.19) was higher than that in GES-1(1.00±0.11),the difference was statistically significant,t=12.15,P<0.001.After plasmid transfection,the expression level of SFRP4in Scr group was 0.95±0.10,while that in SiSFRP4group was 0.27±0.06,and the difference was statistically significant,t=10.10,P<0.001.The number of cells passing through Matrigel membrane of Transwell chamber in SiSFRP4group(0.41±0.10)was lower than that in Scr group(1.00±0.12),and the difference was statistically significant,t=6.542,P=0.003.After transfection for 24and 48h,the cell mobility of SiSFRP4group was 0.42±0.13and 0.41±0.08,the Scr group was 1.00±0.17and 1.00±0.08,and the difference was statistically significant(t values were 4.693and 8.851,Pvalues were 0.009and<0.001).After transfection for 24h,E-cadherin expression in SiSFRP4group(2.35±0.15)was higher than that in Scr group(1.00±0.22),the difference was statistically significant,t=8.782,P <0.001.Expression of Vimentin in SiSFRP4group(0.31±0.14)was lower than that in Scr group(1.00±0.27),and the difference was statistically significant,t=3.930,P=0.017.The expression of MMP-2and MMP-9in SiSFRP4group was0.58±0.09and 0.66±0.08,which was lower than that in Scr group(1.00±0.12,1.00±0.10),and the difference was statistically significant(t values were 4.850and 4.599,P values were 0.008and 0.010).Conclusion SFRP4is highly expressed in gastric cancer cell and can promote the migration and invasion process of gastric cancer cell through EMT pathway and up-regulation of MMPs.