Feng Yangyang,Chen Jie,Mu Ling,Su Linqian,Yi Yongfen.Effect of GM130 gene silencing on proliferation, invasion and migration in human gastric carcinoma cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(12):1217-1221.
Feng Yangyang,Chen Jie,Mu Ling,Su Linqian,Yi Yongfen.Effect of GM130 gene silencing on proliferation, invasion and migration in human gastric carcinoma cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(12):1217-1221.DOI:
Effect of GM130 gene silencing on proliferation, invasion and migration in human gastric carcinoma cells
Western blot和RT-PCR筛选出高表达的GM130细胞株MKN-45、SGC-7901,设计并化学合成、转染、筛选出针对GM130的小干扰RNA片段。通过MTT、Transwell、Matrigel侵袭实验,观测下调GM130后对胃癌细胞株的生物学行为影响,Western blot检测细胞中MMP-2、MMP-9蛋白的变化。 结果 Western blot和RT-PCR检测结果显示,在MKN-45、SGC-7901细胞中GM130蛋白和mRNA水平呈现高表达水平。Western blot和qRT-PCR结果显示在低分化胃癌MKN-45细胞中,GM130-siRNA-519转染组GM130基因的表达水平显著抑制(P0.05)。与对照组相比,抑制转染组细胞的增殖、迁移能力明显下降,穿膜细胞数明显减少,MMP-2、MMP-9的蛋白表达水平显著降低(P0.05)。 结论 抑制GM130基因的表达下调可显著降低胃癌细胞的增殖和体外侵袭转移能力。
Abstract
Objective To explore the expression of Golgi matrix protein 130 (GM130) in different differentiated human gastric carcinoma cell lines and investigate the effect of small interference RNA-mediated GM130 gene silencing on the cell proliferation
invasion and migration in vitro. Methods Three different differentiated gastric carcinoma cell lines (MKN-28
SGC-7901 and MKN-45) were cultured in vitro. Western blot analysis and RT-PCR were carried out to screen MKN-45 cells and SGC-7901cells with high GM130 expression. GM130-targeting siRNA was designed
chemically synthesized and transferred into MKN-45 cells. The effects of GM130 gene silencing on the proliferation
migration and invasion of MKN-45 cells were analyzed by MTT assay
Transwell migration assay and Matrigel invasion assay
respectively. The expression of matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP9) were also detected by Western blotting. Results MKN-45 cells and SGC-7901 cells had high expression of GM130 at mRNA and protein levels. Western blotting and qRT-PCR showed that GM130-siRNA-519 significantly decreased the GM130 expression in the low differentiated MKN-45cells (P0.05). The cell proliferation
migration and invasion of MKN-45 cells transfected with GM130-siRNA were suppressed (P0.05)
and the protein levels of MMP-2 and MMP-9 were significantly decreased (P0.05). Conclusion Small interference RNA-mediated GM130 gene silencing can significantly inhibit the proliferation
migration and invasion in human gastric cancer cell line MKN-45.