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抑制GM130表达对胃癌细胞增殖、侵袭及迁移能力的影响

, PP. 1217-1221

Keywords: GM,RNA干扰,胃肿瘤,增殖,侵袭,迁移

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Abstract:

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),Westernblot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,Westernblot检测细胞中MMP-2、MMP-9蛋白的变化。结果Westernblot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。Westernblot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。

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