title: Regulation of the extrinsic apoptotic pathway by microRNA-21 in alcoholic liver injury.
abstract: IL-6/Stat3 is associated with the regulation of transcription of key cellular regulatory genes (microRNAs) during different types of liver injury. This study evaluated the role of IL-6/Stat3 in regulating miRNA and miR-21 in alcoholic liver disease. By microarray, we identified that ethanol feeding significantly up-regulated 0.8% of known microRNAs in mouse liver compared with controls, including miR-21. Similarly, the treatment of normal human hepatocytes (N-Heps) and hepatic stellate cells (HSCs) with ethanol and IL-6 significantly increased miR-21 expression. Overexpression of miR-21 decreased ethanol-induced apoptosis in both N-Heps and HSCs. The expression level of miR-21 was significantly increased after Stat3 activation in N-Heps and HSCs, in support of the concept that the 5'-promoter region of miR-21 is regulated by Stat3. Using real time PCR, we confirmed that miR-21 activation is associated with ethanol-linked Stat3 binding of the miR-21 promoter. A combination of bioinformatics, PCR array, dual-luciferase reporter assay, and Western blot analysis revealed that Fas ligand (TNF superfamily, member 6) (FASLG) and death receptor 5 (DR5) are the direct targets of miR-21. Furthermore, inhibition of miR-21 by specific Vivo-Morpholino and knock-out of IL-6 in ethanol-treated mice also increased the expression of DR5 and FASLG in vivo during alcoholic liver injury. The identification of miR-21 as an important regulator of hepatic cell survival, transformation, and remodeling in vitro, as well as its upstream modulators and downstream targets, will provide insight into the involvement of altered miRNA expression in contributing to alcoholic liver disease progression and testing novel therapeutic approaches for human alcoholic liver diseases.
authors.affiliation: From the Research, Central Texas Veterans Health Care System and the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504.
authors.forename: Heather
authors.initials: H
authors.lastname: Francis
authors.affiliation: From the Research, Central Texas Veterans Health Care System and the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504.
authors.forename: Kelly
authors.initials: K
authors.lastname: McDaniel
authors.affiliation: the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504.
authors.forename: Yuyan
authors.initials: Y
authors.lastname: Han
authors.affiliation: the Department of Experimental Therapeutics, Division of Cancer Medicine, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030.
authors.forename: Xiuping
authors.initials: X
authors.lastname: Liu
authors.affiliation: From the Research, Central Texas Veterans Health Care System and the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504.
authors.forename: Lindsey
authors.initials: L
authors.lastname: Kennedy
authors.affiliation: the Department of Hepatobiliary Surgery, Shengjing Hospital, China Medical University, Shenyang 100004, China, and.
authors.forename: Fuquan
authors.initials: F
authors.lastname: Yang
authors.affiliation: the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504.
authors.forename: Jennifer
authors.initials: J
authors.lastname: McCarra
authors.affiliation: the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504.
authors.forename: Tianhao
authors.initials: T
authors.lastname: Zhou
authors.affiliation: From the Research, Central Texas Veterans Health Care System and the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504.
authors.forename: Shannon
authors.initials: S
authors.lastname: Glaser
authors.affiliation: the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504.
authors.forename: Julie
authors.initials: J
authors.lastname: Venter
authors.affiliation: the Department of Hepatobiliary Surgery, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China.
authors.forename: Li
authors.initials: L
authors.lastname: Huang
authors.affiliation: the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504.
authors.forename: Phillip
authors.initials: P
authors.lastname: Levine
authors.affiliation: the Department of Hepatobiliary Surgery, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China.
authors.forename: Jia-Ming
authors.initials: JM
authors.lastname: Lai
authors.affiliation: the Department of Experimental Therapeutics, Division of Cancer Medicine, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030.
authors.forename: Chang-Gong
authors.initials: CG
authors.lastname: Liu
authors.affiliation: From the Research, Central Texas Veterans Health Care System and the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504, galpini@tamu.edu.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Alpini
authors.affiliation: From the Research, Central Texas Veterans Health Care System and the Department of Medicine and Scott & White Digestive Disease Research Center, Texas A&M Health Science Center College of Medicine and Scott & White Hospital, Temple, Texas 76504, fmeng@tamu.edu.
authors.forename: Fanyin
authors.initials: F
authors.lastname: Meng
mesh_terms: D000818:Animals; D017209:Apoptosis; D055166:Hepatic Stellate Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008108:Liver Diseases, Alcoholic / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D015854:Up-Regulation
chemical_list: D015850:Interleukin-6; C524940:MIRN21 microRNA, human; C531950:MIRN21 microRNA, mouse; D035683:MicroRNAs; D050796:STAT3 Transcription Factor
keywords: Alcohol; Alcoholic Liver Diseases; DR5; FASLG; Interleukin 6 (IL-6); Liver Injury; MicroRNA (miRNA); STAT3