ARHI真核表达质粒对胃癌恶性表型的影响

The Effect of Eukaryotic Expression Plasmid ARHI on Gastric Cancer

目的观察外源ARHI在胃癌细胞中过表达对胃癌细胞增殖、迁移以及侵袭的影响。方法构建pEGFP-ARHI表达载体,并通过lipofectamineTM2000将其转入ARHI低表达的胃癌细胞系MKN-28中作为实验组,空载质粒pEGFP-N1转入MKN-28作为空载组,未处理MKN-28细胞作为未处理组。通过荧光显微镜及Western blot检测外源ARHI的表达,采用增殖实验、损伤刮擦实验、体外侵袭实验(Transwell小室)分别检测过表达ARHI胃癌细胞MKN-28的增殖能力、侵袭和迁移能力的变化。结果 ARHI在胃癌细胞系MKN-28中表达较低,成功构建重组真核表达质粒pEGFP-ARHI并成功转染MKN-28细胞后,CCK8法检测显示,相对于空载组和未处理组,转染pEGFP-ARHI的实验组细胞增殖变慢(P<0.05),Transwell小室实验显示实验组细胞侵袭能力减弱(P<0.05),细胞划痕实验表明实验组细胞迁移能力减弱(P<0.05)。结论在ARHI低表达的胃癌细胞系MKN-28中表达重组质粒pEGFP-ARHI能够抑制细胞的增殖、侵袭和迁移能力。

Objective To study the effect of expressed aplasia ras homolog member I （ARHI） on the malignant biological behaviors of gastric cancer including the proliferation, migration and invasion of the cell. Methods The eukaryotic expression plasmid of ARHI was constructed and transfected into MKN-28 cell with lipofectamineXM2000 as pEGFP-ARHI group, transfected with pEGFP-N1 as pEGFP-N1 group, and untreated MKN-28 as control group. The expression of ARHI was detected by Western blotting and fluorescence microscope. CCK-8 assay was used to analyze the cell proliferation, the wound-healing assay and transwell assay were performed to investigate the effects on migration and invasion. Results Compared with the pEGFP-N1 group and＇ control group, proliferation, invasion and migration of the pEGFP-ARHI group were depressed （P〈0.05）. Conclusion Recombination eukaryotic expression pEGFP-ARHI could partially reverse the malignant phenotypes of gastric cancer cell MKN-28.