基础研究(Basic Research)

GPSM2过表达对人胰腺癌细胞迁移能力的影响

Published at: 2017年第26卷第3期

魏彪 1 , 张建新 1 , 崔磊 1 , 瞿建国 1 , 钱小宝 1 , 党胜春 1
1 江苏大学附属医院 普通外科,江苏 镇江 212001
通讯作者 胜春 党 Email: dscgu@163.com
DOI: 10.3978/j.issn.1005-6947.10.3978/j.issn.1005-6947.2017.03.007
基金:
江苏省自然科学基金资助项目 BK2012704
江苏省博士后科研计划基金资助项目 1302096B

摘要

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-Tag 3B-GPSM2)并鉴定,将人胰腺癌MIA-PaCa-2细胞分别转染pCMV-Tag 3B-GPSM2(GPSM2转染组)或pCMV-Tag 3B空载体(阴性对照组),以无处理的MIA-PaCa-2细胞为空白对照,用RT-PCR检测各组细胞GPSM2 mRNA表达;Western blot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。


Effects of GPSM2 over-expression on migration ability of human pancreatic cancer cells

Abstract

Objective: To construct a stable pancreatic cancer cell line with high expression of the G-protein signaling modulator 2 (GPSM2), for investigating the relationship between GPSM2 and migration ability of human pancreatic cancer cells. Methods: The plasmids over-expressing GPSM2 gene (pCMV-Tag 3B-GPSM2) were constructed and then were identified. Human pancreatic cancer MIA-PaCa-2 cells were transfected with pCMV-Tag 3B-GPSM2 (GPSM2 transfection group) or empty pCMV-Tag 3B vectors (negative control group), with untreated MIA-PaCa-2 cells as blank control. In each group of cells, the GPSM2 mRNA expressions were measured by RT-PCR, the protein expressions of GPSM2 and β-catenin were determined by Western blot analysis, and the migration abilities were tested by Transwell assay, respectively. Results: The recombinant cell line with stable high GPSM2 expression was successfully constructed. In GPSM2 transfection group compared with blank control group, the GPSM2 mRNA expression was significantly up-regulated, with a 73.3-fold up-regulation, the protein expression levels of GPSM2 and β-catenin were significantly elevated, and the number of migrated cells was significantly increased (all P<0.05). In addition, a positive linear relationship existed between GPSM2 and β-catenin expressions in pancreatic cancer cells (P<0.05). There was no statistical difference in any of the indexes between negative control group and blank control group (all P>0.01). Conclusion: Up-regulating GPSM2 expression in pancreatic cancer cells can increase the migration ability of pancreatic cancer cells, which may be associated with increased β-catenin protein expression.


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引用

引用本文: 彪 魏, 建新 张, 磊 崔, 建国 瞿, 小宝 钱, 胜春 党. GPSM2过表达对人胰腺癌细胞迁移能力的影响[J]. 中国普通外科杂志, 2017, 26(3): 311-316.
Cite this article as: WEI Biao, ZHANG Jianxin, CUI Lei, QU Jian'guo, QIAN Xiaobao, DANG Shengchun . Effects of GPSM2 over-expression on migration ability of human pancreatic cancer cells [J]. Chin J Gen Surg, 2017, 26(3): 311-316.