title: [Establishment and application of a high-throughput drug screening model based on COL1A1 promoter for anti-liver fibrosis].
abstract: For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
authors.forename: Shuang-Shuang
authors.initials: SS
authors.lastname: Zhao
authors.forename: Ju-Xian
authors.initials: JX
authors.lastname: Wang
authors.forename: Yu-Cheng
authors.initials: YC
authors.lastname: Wang
authors.forename: Rong-Guang
authors.initials: RG
authors.lastname: Shao
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: He
mesh_terms: D024042:Collagen Type I / Q000235:genetics*; D000090602:Collagen Type I, alpha 1 Chain; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D017930:Genes, Reporter; D055166:Hepatic Stellate Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008103:Liver Cirrhosis / Q000188:drug therapy*; D008156:Luciferases; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
chemical_list: D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D012333:RNA, Messenger; D053773:Transforming Growth Factor beta1; D008156:Luciferases