title: BCL2 promotor methylation and miR-15a/16-1 upregulation is associated with sanguinarine-induced apoptotic death in rat HSC-T6 cells.
abstract: Previous studies show that several pathways are involved in sanguinarine-induced apoptotic cell death, including AKT downregulation, inhibition of NF-kB activation, mediation of ROS production, downregulation of anti-apoptosis proteins XIAP and cIAP-1, upregulation of BAX, and downregulation of BCL2. In this study, we found out that the quenching of ROS generation by N-acetyl-l-cysteine (NAC), a scavenger of ROS, reversed sanguinarine-induced apoptosis effects, also we found out that sanguinarine-induced rat hepatic stellate T6 cells (HSC-T6 cells) apoptosis was correlated with the generation of increased ROS, which was followed by the activation of caspase-8 (-3, -6, and -9), and the decreasing in the miltochondrial membrane potential (MMP) and the down-regulation of anti-apoptotic protein Bcl-2. It is not clear whether BCL2's downregulation relates to its promoter methylation and miR-15a/16-1 expression which can bind to BCL2 3'-UTR (un-translation reagon). We showed that sanguinarine-induced down regulation of BCL2 was associated with the increased methylation rate of BCL2 promotor district and the increased expression of miR-15a/16-1. HSC-T6 cells treatment with 5-Aza-2'-deoxycytidine (5'-Aza-CdR) impeded sanguinarine-induced BCL2 promotor district methylation and recovered BCL2's expression. Over expression of BCL2 using pEGFP-N1 vector decreased sanguinarine-induced HSC-T6 cells apoptotic death significantly but not completely. These observations clearly showed that BCL2 down regulation was associated with its promoter methylation and miR-15a/16-1 upregulation in sanguinarine-induced Rat HSC-T6 cells.
authors.affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan 410128, China.
authors.forename: Da-Sheng
authors.initials: DS
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan 410128, China.
authors.forename: Yuan-Yi
authors.initials: YY
authors.lastname: Li
authors.affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan 410128, China.
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Chen
authors.affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan 410128, China.
authors.forename: Yu-Juan
authors.initials: YJ
authors.lastname: Li
authors.affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan 410128, China.
authors.forename: Zhao-Ying
authors.initials: ZY
authors.lastname: Liu
authors.affiliation: Department of Pharmaceutics, Hunan University of Traditional Chinese Medicine, Xueshi Road 300, Changsha, Hunan 410208, China.
authors.forename: Wen-Jian
authors.initials: WJ
authors.lastname: Xie
authors.affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan 410128, China; Medicine Resources for Livestock and Innovative Engineering Research Center in Veterinary Medicine, China. Electronic address: sunzhiliang1965@aliyun.com.
authors.forename: Zhi-Liang
authors.initials: ZL
authors.lastname: Sun
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D053119:Benzophenanthridines / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D000077209:Decitabine; D015536:Down-Regulation; D007546:Isoquinolines / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008745:Methylation / Q000187:drug effects; D035683:MicroRNAs / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D053119:Benzophenanthridines; D007546:Isoquinolines; C547573:MIRN16 microRNA, rat; D035683:MicroRNAs; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; C534110:mirn15 microRNA, rat; D000077209:Decitabine; C005705:sanguinarine; D001374:Azacitidine
keywords: Apoptosis; Methylation; MiR-15a/16-1; ROS; Sanguinarine