The Researth Of Differential Genes Expression Profiles Of Human Fibroblasts Reprogrammed By Piwil2

Objective: To study differential genes and the main signaling pathways of the human fibroblasts,which is changed to cancer stem cell-like cells by reprogrammed with piwil2,using gene microarray,and to study the meaning of differential gene which is high expressed in the gene expression profiling in Wilms' tumor.Methods: The foreskin fibroblasts was reprogrammed by lentiviral vector containing piwil2-gfp gene or gfp gene(FB-piwil2: being reprogrammed by piwil2-gfp;FB-GFP: being reprogrammed by gfp;FB: fibroblasts)to built a cancer stem cell-like cell line.Its genome expression changes were profiled with a NimbleGen human gene expression microarray followed pathway analysis.The genes in the Valine,leuc ine and isoleucine degradation singaling pathway which is up-regulated in the Microarray result were verified with q-PCR.Then we measured the expression of ulbp2 which is high expressed in the gene expression profiling in Wilms' tumor using immunohistochemistry and studied the correlation of prognosis of Wilms' tumor and the clinicopathologic feature,such as the expression of ULBP2 protein.Results: Compared with the FB,a total of 1 490 identified genes are up-regulated and 1 484 identified genes are down-regulated in FB-piwil2.These altered genes are involved in some pathways,such as Valine,leuc ine and isoleuc ine degradation signaling pathway and.Compared with the FB-GFP,a total of 3 765 identified genes are up-regulated and 1 988 identified genes are down-regulated in FB-piwil2.These altered genes are involved in some pathways,such as Valine,leuc ine and isoleuc ine degradation s ignaling pathway.Compared with the FB,a total of 2 095 identified genes are up-regulated and 4 014 identified genes are down-regulated in FB-GFP.These altered genes are involved in some pathways,such as Osteoclast differentiation signaling pathway.The genes(acad8,acadm,aldh3a2,bcat1,hibch,hmgcs1,mccc2)in Valine,leuc ine and isoleuc ine degradation signaling pathway were verified using q-PCR,which reveals that microarray analys is results are accurate.ULBP2 protein expression,different pathological type,different clinical stage have significant difference(P < 0.05)in assessing the prognosis of Wilms' tumor.The sensitivity,the specificity,the positive predictive value and the negative predictive value of ULBP2 protein high expression in the diagnosis of Wilms' tumor with poor prognosis are 100%,78.87%,85.7%,and 100%.Conclusion: The human fibroblasts reprogrammed by piwil2 appears differential gene expression,which is in connection with Valine,leuc ine and isoleucine degradation and Natural killer cell mediated cytotoxicity.The differential genes expression profiles can provide clues for the basic study of Wilms' tumor.The ULBP2 protein high expression has high clinical value in assessing the prognosis of Wilms' tumor.