抑制GM130表达对胃癌细胞增殖侵袭及迁移能力-第三军医大学学报.DOCVIP

抑制GM130表达对胃癌细胞增殖、侵袭及迁移能力的影响 冯阳阳，陈 婕，牟 玲，苏琳茜，易永芬 （400016重庆，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） [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 体外培养三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﹤0.05MMP-2、MMP-9的蛋白表达水平显著降低（P﹤0.05GM130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。 [关键词] GM130；RNA干扰；胃肿瘤；增殖；侵袭；迁移 [中图法分类号] [文献标志码] A Effects of GM130 gene suppression on proliferation, invasion and migration of human gastric carcinoma cell Feng Yangyang, Chen Jie, Mu Ling, Su Linqian, Yi Yongfen (Department of Pathology，Molecular Medicine and Cancer Research Center，Chongqing Medical University，Chongqing，400016，China) [Abstract ]Objective To explore the expression of GM30 in the differentiation of gastric carcinoma cell lines and investigate the effect of small interference RNA-mediated silcencing of the GM130 gene on cell proliferation, invasion and migration of human gastric carcinoma MKN-45 cells in vitro. Methods Three different differentiated gastric carcinoma cell lines (MKN-28, SGC-7901 and MKN-45) were cultured in vitro. Western blot and RT-PCR analyses were carried out in MKN-45 cells and SGC-7901cells, which had high GM130 expression levels.The designed and chemically synthesized siRNA targeting the GM130 gene was transfected into MKN-45 cells. The effect of GM130 knockdown on MKN-45 cells proliferation was determined by MTT assay, cells migration was analyzed by Transwell migration assay, the invasion of cells was determined by Matrigel invasion assay. The expression of MMP-2 and -9 was also detected at the protein level. Results MKN-45 and SGC-7901 cells showed the higher expression of GM130 at protein levels, so We