Gene Expression Of Human IntrahepaticCholangiocarcinoma RBE(NCAM~+c-Kit~+)Cell Line

Objective:To analyze the differentially expressed genes between NCAM+c-Kit+RBE and NCAM-c-Kit-RBE of intrahepatic cholangiocarcinoma(ICC) cell line, then classified the differentially expressed genes,and screen out the differentially expressed genes that related to stem cell signaling pathways. So then to prove the characteristic of hepatic progenitor cell of NCAM+c-Kit+RBE futher.In order to reveal the underlying molecular mechanism of intrahepatic cholangiocarcinoma’s occurrence and development,which may provide new experimental bases for target therapy of intrahepatic cholangiocarcinoma.Methods:Magnetic activated cell sorting was used to isolate NCAM+c-Kit+and NCAM-c-Kitsubset cells, and then Agilent Whole Human Genome Microarray Kit was used to test the difference of gene expression between NCAM+c-Kit+and NCAM-c-Kit-subset cells,and get the whole differentially expressed genes. Then the difference of gene expression related to stem cell signaling pathways was analyzed by SAS system. The result of the microarray was futher confirmed by RT-PCR.Results:1.The total differentially expressed genes can be found through gene microarray were7270[foldchange(fc)≥2 or fc≤0.5]. Compared to NCAM-c-Kit-RBE, 3572 genes were upregulated while 3698 genes were downregulated.2.SAS analysis system screened out differences of gene expression related to stem cell signaling pathways were 421(fc≥2 or fc≤0.5), among which 231 genes were upregulated while 190 genes were downregulated compared to NCAM-c-Kit-RBE.3.The results in hedgehog signaling pathway showed by RT-PCR were:compared to NCAM-c-Kit-RBE,Shh had no statistically significant difference(t=-2.183,P>0.05);Ptch1had low expression(t=2.76,P<0.05);Smo had high expression(t=17.17,P<0.001);Gli2 had high expression too(t=10.59,P<0.001).Conclusion:1.High-flux microarray can be used to screen out lots of differentially expressed genesbetween NCAM+c-Kit+and NCAM-c-Kit-RBE cells.2.Compared to NCAM-c-Kit-RBE,NCAM+c-Kit+RBE expressed abnomal activation hedgehog signaling pathway more.