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目的:构建G蛋白信号调节蛋白2(GPSM2)稳定高表达的胰腺癌细胞株,探讨GPSM2与人胰腺癌细胞迁移能力的关系。方法:构建GPSM2基因过表达质粒(pCMV-Tag3B-GPSM2)并鉴定,将人胰腺癌MIA-PaCa-2细胞分别转染pCMV-Tag3B-GPSM2(GPSM2转染组)或p CMV-Tag3B空载体(阴性对照组),以无处理的MIA-Pa Ca-2细胞为空白对照,用RT-PCR检测各组细胞GPSM2 mRNA表达;Westernblot检测各组细胞GPSM2、β-连环蛋白(β-catenin)的表达;用Transwell实验检测各组胰腺癌细胞迁移能力。结果:成功构建了GPSM2稳定高表达的重组细胞株。与空白对照组比较,GPSM2转染组细胞GPSM2 mRNA表达量明显上调,达前者73.3倍、GPSM2、β-catenin蛋白表达量明显升高、迁移细胞计数明显增加(均P<0.05)。此外,胰腺癌细胞中GPSM2与β-catenin的表达水平呈明显的正向线性关系(P<0.05)。阴性对照组与空白对照组间各指标的差异均无统计学意义(均P>0.05)。结论:上调胰腺癌细胞中GPSM2的表达能增加胰腺癌细胞的迁移能力,该作用可能与β-catenin蛋白表达升高有关。
OBJECTIVE: To construct a stable and stable pancreatic cancer cell line expressing G protein signaling 2 (GPSM2) and investigate the relationship between GPSM2 and human pancreatic cancer cell migration. METHODS: The human CMM2 gene overexpression plasmid (pCMV-Tag3B-GPSM2) was constructed and transfected into pCMV-Tag3B-GPSM2 (GPSM2 transfection group) or pCMV-Tag3B empty vector Negative control group). The untreated MIA-Pa Ca-2 cells were used as a blank control, and the expression of GPSM2 mRNA was detected by RT-PCR. The expression of GPSM2 and β-catenin in each group was detected by Western blot Transwell assay was used to detect the migration of pancreatic cancer cells in each group. Results: The recombinant cell line with stable and high expression of GPSM2 was successfully constructed. Compared with the blank control group, the expression of GPSM2 mRNA in GPSM2 transfection group was significantly increased up to 73.3-fold, and the expression of GPSM2 and β-catenin protein was significantly increased (P <0.05). In addition, pancreatic cancer cells in the expression of GPSM2 and β-catenin showed a significant positive linear relationship (P <0.05). There was no significant difference between the negative control group and the blank control group (all P> 0.05). Conclusion: Upregulation of GPSM2 in pancreatic cancer cells can increase the migration ability of pancreatic cancer cells, which may be related to the increase of β-catenin protein expression.