抑制GM130表达对胃癌细胞增殖、侵袭及迁移能力的影响  

Effect of GM130 gene silencing on proliferation,invasion and migration in human gastric carcinoma cells

作　　者：冯阳阳[1] 陈婕[1] 牟玲[1] 苏琳茜 易永芬[1] 

机构地区：[1]重庆医科大学基础医学院病理学教研室分子医学与肿瘤研究中心,重庆400016

出　　处：《第三军医大学学报》2015年第12期1217-1221,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金(30672431);高等学校博士学科点专项科研基金(20060631006)~~

摘　　要：目的分析GM130在不同分化人胃癌细胞系的差异表达,利用小干扰RNA技术来沉默GM130基因,研究其对胃癌细胞生物学行为的影响。方法体外培养3株不同分化程度胃癌细胞系(高分化MKN-28、中分化SGC-7901、低分化MKN-45),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(P<0.05)。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(P<0.05)。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。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-7901 cells 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（ P 〈 0. 05）. The cell proliferation,migration and invasion of MKN-45 cells transfected with GM130-siRNA were suppressed（ P 〈 0. 05）,and the protein levels of MMP-2and MMP-9 were significantly decreased（ P 〈 0. 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.

关 键 词：GM130 RNA干扰 胃肿瘤 增殖 侵袭 迁移 

分 类 号：R394.3[医药卫生—医学遗传学] R73-362[医药卫生—基础医学]