p115基因沉默对人胃癌BGC-823细胞迁移及侵袭能力的影响及其机制

Effect of p115 gene silence on migration and invasion abilities of human gastric cancer BGC-823 cells and relevant mechanism

目的探讨高尔基体囊泡转运蛋白(Golgi-vesicular transport protein)p115基因沉默对人胃癌BGC-823细胞侵袭能力的影响及其机制。方法采用脂质体法将质粒pGPU6/GFP/Neo/p115(p115 shRNA)和阴性对照质粒(shNC)分别转染至BGC-823细胞中,并设空白对照组。转染后48 h,采用RT-PCR、Westernblot和细胞免疫荧光法检测各组细胞中p115基因及蛋白表达的变化,及其对巨噬细胞游走抑制因子(Macrophage migration inhibitory factor,MIF)、基质金属蛋白酶-2(Matrix metallopro-teinase-2,MMP-2)、MMP-9基因和蛋白表达的调节;细胞划痕及Transwell小室试验检测细胞迁移及侵袭能力的变化。结果p115 shRNA组细胞中p115、MIF、MMP-2、MMP-9在基因和蛋白水平的表达均较shNC组和空白对照组明显下降(P<0.01);p115shRNA组细胞的划痕愈合能力明显低于两个对照组(P<0.01);p115 shRNA组的穿膜细胞数明显低于两个对照组(P<0.01)。结论在BGC-823细胞中,抑制P115的表达后,可能通过下调MIF、MMP-2、MMP-9的表达,抑制肿瘤细胞的侵袭运动。

Objective To investigate the effect of Golgi-vesicular transport protein pl15 gene silence on migration and invasion abilities of human gastric cancer BGC-823 cells as well as the relevant mechanism. Methods BGC-823 cells transfected with plasmid pGPU6/GFP/Neo/p]15 （p115 shRNA） and negative control plasmid （shNC） in mediation of liposome respectively, using those untransfected as blank control. Forty-eight hours after transfection, the expressions of pllS, macrophage migration inhibitory factor （MIF）, matrix metalloproteinase-2 （MMP-2） and MMP-9 genes and proteins in various groups were determined by RT-PCR, Western blot and IFA, while the invasion ability of cells by wound healing test and Transwell test. Results Compared with those in shNC and blank control groups, the expression levels ofpllS, MIF, MMP-2 and MMP-9 gene and protein as well as the healing ability and the number of transmembrane cells in pl15 shRNA group decreased significantly （each P 〈 O. O1 ）. Conclusion Inhibition of PlI5 expression in BGC-823 cells inhibited the invasion ability of tumor cells by down-regulating the expressions of MIF, MMP-2 and MMP-9 genes.