慢病毒介导的Annexin A2基因沉默对胃癌AGS细胞基因表达谱的影响

The effect of lentivirus-mediated Annexin A2 silence on gene expression profiles of AGS gastric cancer cells

目的:探讨膜联蛋白A2(Annexin A2,ANXA2)基因沉默对AGS胃癌细胞基因表达谱的改变。方法:通过慢病毒干扰AGS细胞ANXA2的表达,用人基因表达谱芯片筛选ANXA2调控的下游基因。结果:通过基因表达谱芯片筛选出ANXA2沉默后的共同差异表达基因100个,其中58个上调的基因,42个下调的基因。根据KEGG与BIOCARTA中所有Pathway的基因信息,将差异基因进行富集分析。筛选到的ANXA2调控的重要相关基因包括FGA、FGB、SERPINB2、CD55、PLAUR、MET、RAP1A和ETS1。Western blot进一步验证结果显示,沉默ANXA2后,AGS细胞中RAP1A表达水平降低,MAP2K1的蛋白表达亦降低。结论:p GC-LVANXA2 siRNA可以有效地下调胃癌AGS细胞ANXA2基因的表达。RAP1A和MAP2K1可能是AGS胃癌细胞中ANXA2参与MAPK信号通路的潜在下游调控基因。

Objective：To investigate whether the Annexin A2 gene silencing changes gene expression profiles of AGS gastric cancer cells. Methods：Lentivirus was used to interfer the ANXA2 expression in AGS cells,and the gene expression profile was carried out to screen the downstream genes of ANXA2. Results：ANXA2 expression level was significantly declined in AGS gastric cancer cells transfected with pGC - LV - ANXA2 siRNA. Using gene expression microarray,100 differentially expressed genes were detected,including 58 up - regulated human genes and 42 down -regulated human genes. According to all Pathway genetic information in KEGG and BIOCARTA,we did the enrich-ment analysis of the difference of gene. The important ANXA2 related genes including FGA,FGB,SERPINB2,CD55, PLAUR,MET,RAP1A and ETS1 were screened. Western blot showed that,after silencing of ANXA2,RAP1A and MAP2K1 protein expression decreased in AGS cells. Conclusion：PGC - LV - ANXA2 siRNA could effectively down- regulate the expression of human ANXA2 gene in human gastric AGS cancer cell. RAP1A and MAP2K1 may be po-tential downstream regulating genes of ANXA2 taking part in MAPK signaling pathway in AGS gastric cancer cells.