基因芯片检测c-Met表达对结直肠癌侵袭和转移的机制研究

Study on the mechanism of c-Met expression on invasion and metastasis of colorectal cancer by gene microarray

目的研究c-Met基因下调后对结直肠癌侵袭和转移的影响机制。方法在SW480细胞中敲低c-Met基因,通过基因芯片筛选差异基因,对差异基因进行功能聚类分析,通过IPA分析细胞信号通路和相关的差异基因相互作用网络以及相关基因与上游调控分子间的作用关系。选取被抑制的信号通路中的相关基因进行qPCR验证。结果敲低c-Met后,SW480细胞中上调基因数目399个,下调基因数目286个。聚类分析热图提示:c-Met敲低后,对SW480细胞的基因表达水平产生了明显影响;IPA通路分析表明:HGF信号通路被抑制,且c-Met敲低后,IPA相互作用网络提示:HGF通路中的AKT2、PIK3CA及MAP2K4下调;qPCR验证上述基因亦下调,与芯片结果相符。结论 c-Met可能通过对HGF通路中AKT2、PIK3CA及MAP2K4的调控影响了结直肠癌的侵袭、转移。

Objective To study the mechanism of the effect on invasion and metastasis of colorectal cancer by down-regulating c-Met gene.Methods c-Met genes were knocked down in SW480 cells,differential genes were screened by gene chip,functional cluster analysis of differential genes was carried out,and IPA was used to analyze the interaction network of cell signal pathway and related differential genes,as well as the ralationship between related genes and upstream regulatory molecules.The related genes in the suppressed signal pathway were selected for qPCR verification.Results After knockdown of c-Met,the number of up-regulated genes and down-regulated genes in SW480 cells was 399 and 286,respectively.Cluster analysis showed that c-Met knockdown had a great effect on the gene expression level of SW480 cells,IPA pathway analysis showed that HGF signaling pathway was suppressed,and after c-Met knockdown,IPA interaction network suggested that AKT2,PIK3CA,and MAP2K4 in HGF pathway were down-regulated,and qPCR verified that the above genes were also down-regulated,which was consistent with the results of microarray.Conclusion c-Met may affect the invasion and metastasis of colorectal cancer through the regulation of AKT2,PIK3CA,and MAP2K4 in HGF pathway.