微球蛋白1对胃癌细胞侵袭和迁移的影响及其机制

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Lipo3000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。

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

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 N-cadherin 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.