微球蛋白1对胃癌细胞侵袭和迁移的影响及其机制

Effects of microspherule protein 1 on invasion and migration of gastric cancer cells and their mechanisms

目的:探讨微球蛋白1 (MCRS1)在胃癌细胞中的过表达对胃癌细胞侵袭和迁移的影响,并阐明其可能的作用机制。方法:选择胃癌BGC-823细胞、SGC-7901细胞和正常胃黏膜上皮GES-1细胞进行培养,采用Western blotting法检测MCRS1在3种细胞中表达情况,并选择MCRS1蛋白表达低的胃癌BGC-823细胞进行后续实验。构建MCRS1重组质粒,选取处于对数生长期的胃癌BGC-823细胞,设立空白组、空载体转染组和MCRS1转染组,利用Lipo 3000将质粒转染入BGC-823细胞,采用Western blotting法检测侵袭相关蛋白上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)及Snail的表达水平,采用细胞划痕实验和Transwell小室实验检测各组胃癌细胞的迁移和侵袭能力。结果:与正常胃黏膜上皮GES-1细胞比较,MCRS1在胃癌BGC-823细胞中表达水平较低(P<0.01),而在胃癌SGC-7901细胞中表达水平较高(P<0.01)。PCR鉴定和测序分析,MCRS1重组质粒构建成功。与空白组和空载体转染组比较,MCRS1转染组MCRS1和E-cadherin蛋白表达水平明显升高(P<0.01),N-cadherin和Snail蛋白表达水平明显降低(P<0.01),细胞迁移率明显降低(P<0.01),侵袭细胞数明显减少(P<0.01)。结论:过表达MCRS1能抑制胃癌BGC-823细胞的迁移和侵袭,其机制可能与E-cadherin蛋白表达增加、N-cadherin和Snail蛋白表达降低有关。

Objective:To investigate the effects of overexpression of microspherule protein 1(MCRS1)in the gastric cancer cells on the invasion and migration of gastric cancer cells,and to elucidate their possible mechanisms.Methods:The gastric cancer BGC-823 cells,SGC-7901 cells and the normal gastric mucosal epithelial GES-1 cells were cultivated.Western blotting method was used to detect the expressions of MCRS1 in three kinds of cells.The result showed that MCRS1 protein had the lowest expression in the BGC-823 cells,so the gastric cancer BGC-823 cells were selected for next experiments.The recombinant plasmid of MCRS1 was constructed,and the gastric cancer BGC-823 cells in the logarithmic growth phase were selected.Blank group,empty vector transfection group and MCRS1 transfection group were established,and the plasmid was transfected into the BGC-823 cells using Lipo3000.The expression levels of epithelial cadherin(E-cadherin)protein,neuronal cadherin(N-cadherin)protein and Snail protein were detected by Western blotting method.The cell scratch assay and Transwell assay were used to detect the migration and invasion of gastric cancer BGC-823 cells.Results:Compared with the normal gastric epithelial GES-1 cells,the expression level of MCRS1 protein in the gastric cancer BGC-823 cells was decreased(P<0.01),but the expression level of MCRS1 protein in the SGC-7901 cells was increased(P<0.01).The PCR identification and sequencing analysis showed that the MCRS1 recombinant plasmid was successfully constructed.Compared with blank group and empty vector transfection group,the expression levels of MCRS1 protein and E-cadherin protein in MCRS1 transfection group were increased(P<0.01),the expression levels of Ncadherin and Snail proteins were decreased(P<0.01),the cell migration rate was significantly reduced(P<0.01),and the number of invasion cells was significantly decreased(P<0.01).Conclusion:Overexpression of MCRS1 can inhibit the migration and invasion of gastric cancer BGC-823 cells,which may be related to the increased expression of E-cadherin protein and the decreased expressions of N-cadherin and Snail proteins.