Objective: To screen the differentially expressed genes in hepatic stellate cells (HSC) treated with transforming growth factor beta 1 (TGFbeta1) by cDNA microarray technique, and to elucidate the molecular pathogenesis of liver fibrosis involving TGFb1.
Methods: Total RNA was extracted from HSC treated with TGFbeta1 and PBS by trizol and reverse-transcribed to double strand cDNA templates. Transcription of cDNA probe with biotin-labeling was performed, and then the obtained cDNA was hybridized with human cDNA microarray. The results were imaged by an Agilent scanner, and the differentially expressed genes were analyzed with bioinformatics software.
Results: One hundred seventy-seven differentially expressed genes were screened from 13824 targeting genes; 123 genes were up-regulated, including connective tissue growth factor, tubulin epsilon 1, collagen, type V, alpha2, catenin delta 2, cadherin 6, type 2, Smad3, mitogen-activated protein kinase 4, growth factor receptor-bound protein 7 and MAP kinase-interacting serine/threonine kinase 1; 54 genes were down-regulated, including TNF receptor-associated factor 4, interferon regulatory factor 7, interferon inducible protein p78, bone morphogenetic protein 7, matrix gla protein, serine proteinase inhibitor, interferon stimulated gene 2.0 x 10(4), death-associated protein 6, metallothionein 1H and superoxide dismutase 2; in addition, 8 genes with unknown functions were also found.
Conclusion: The differentially expressed genes in HSC treated with TGFbeta1 were successfully screened by cDNA microarray technique. It revealed that the molecular pathogenesis of liver fibrosis involving TGFbeta1 was the result of co-regulation by multiple factors. This information might be of help in searching for new targets in gene therapy.