title: miR-15b and miR-16 induce the apoptosis of rat activated pancreatic stellate cells by targeting Bcl-2 in vitro.
abstract: OBJECTIVE
  Activated pancreatic stellate cells (PSCs) play a pivotal role in the development of pancreatic diseases, especially chronic pancreatitis and pancreatic cancer. MicroRNAs have become a focal point of interest as post-transcriptional regulators of gene expression via their interaction with the 3' untranslated region of target mRNAs, which results in gene silencing. We examined the relative expression of microRNAs (miR-15b and miR-16) and their target gene, Bcl-2, during activation of rat PSCs, and determined their effects on apoptosis of rat PSCs in vitro.
  
  
  METHODS
  miR-15b and miR-16 expression levels were analyzed in quiescent and activated PSCs by stem-loop RT-PCR. In addition, the effects of miR-15b and miR-16 on apoptosis of activated PSCs were investigated by immunofluorescence microscopy with Hoechst 33342 staining, and flow cytometry with annexin-V/propidium (PI) co-labeling. Bcl-2 and Bcl-xl were also analyzed by real-time RT-PCR and Western blotting.
  
  
  RESULTS
  During activation of PSCs, from the quiescent stage to activated stage, miR-15b and miR_16 were downregulated, while Bcl-2 expression was upregulated. Restoring intracellular miRNA levels by miR-15b and miR-16 administration greatly reduced Bcl-2 protein levels, and significantly induced apoptosis in activated PSCs.
  
  
  CONCLUSIONS
  miR-15b and miR-16 could induce apoptosis of rat PSCs by targeting Bcl-2.
authors.affiliation: Department of Gastroenterology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Shen
authors.forename: Rong
authors.initials: R
authors.lastname: Wan
authors.forename: Guoyong
authors.initials: G
authors.lastname: Hu
authors.forename: Lijuan
authors.initials: L
authors.lastname: Yang
authors.forename: Jie
authors.initials: J
authors.lastname: Xiong
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.forename: Jiaqing
authors.initials: J
authors.lastname: Shen
authors.forename: Shanshan
authors.initials: S
authors.lastname: He
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Guo
authors.forename: Jianbo
authors.initials: J
authors.lastname: Ni
authors.forename: Chuanyong
authors.initials: C
authors.lastname: Guo
authors.forename: Xingpeng
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D017209:Apoptosis; D001483:Base Sequence; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D020868:Gene Silencing / Q000187:drug effects; D019254:Genes, bcl-2 / Q000187:drug effects / Q000235:genetics*; D008297:Male; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009696:Nucleic Acids / Q000494:pharmacology; D010179:Pancreas / Q000166:cytology*; D058954:Pancreatic Stellate Cells / Q000187:drug effects / Q000473:pathology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects
chemical_list: C547573:MIRN16 microRNA, rat; D035683:MicroRNAs; C570833:Mirn15 microRNA, mouse; D009696:Nucleic Acids; D019253:Proto-Oncogene Proteins c-bcl-2