title: [microRNA-203 INDUCING DIFFERENTIATION OF HUMAN EPIDERMAL STEM CELLS INTO SWEAT GLAND CELLS IN VITRO].
abstract: OBJECTIVE
  To observe the possibility and mechanism of microRNA (miRNA)-203 inducing the human epidermal stem cells to differentiate into sweat gland cells.
  
  
  METHODS
  Five normal human foreskin tissues were harvested to prepare a single cell suspension by 0.25% trypsin-EDTA digestion method, then the human epidermal stem cells were isolated and cultured by type IV collagen differential adherent method. The cell morphology was observed by inverted phase contrast microscope. The monoclonal antibodies of integrin β1 (ITGB1), cytokeratin19 (CK19), CK1, CK10, CK18, and carcinoembryonic antigen (CEA) were used for identification by immunocytochemical staining. Double stranded mimics of has-miR-203 were transfected into the human epidermal stem cells with Lipofectamine 2000 (experimental group) and the human epidermal stem cells transfected with nonsense miRNA mimics served as control group. The monoclonal antibodies of ITGB1, CK19, CK1, CK10, CK18, and CEA were used for identifying the cells after transfection by immunocytochemical staining; the mRNA relative expressions of miRNA-203, P63, ITGB1, CK19, CK1, CK1O, CK18, and CEA were detected by real-time fluorescence quantitative RT-PCR before transfected and at 72 hours after transfected. The protein relative expressions of P63, ITGB1, CK19, CK1, CK10, CK18, and CEA were detected by Western blot. The mRNA expression of miRNA-203 and the mRNA and protein expressions of P63 were analyzed respectively with Pearson correlation.
  
  
  RESULTS
  The CK19 and ITGB1 were positively expressed before transfection, but CK1, CK10, CK18, and CEA were expressed positively after transfection. The mRNA relative expression of miRNA-203 after transfection in experimental group was significantly higher than that before transfection (P < 0.05). The mRNA and protein relative expressions of CK1, CK10, CK18, and CEA after transfection in experimental group were significantly higher than those before transfection and control group (P < 0.05), while the mRNA and protein relative expressions of P63, CK19, and ITGB1 were significantly lower than those before transfection and control group (P < 0.05). These indicators showed no significant difference between the control group and before transfection (P > 0.05). The expression level of miRNA-203 was negatively correlated with the mRNA and protein relative expressions of P63 before and after transfection, the correlation coefficients before transfection were -0.91 (t = 3.862, P = 0.042) and -0.96 (t = 5.971, P = 0.009) respectively; the correlation coefficients after transfection were -0.92 (t = 4.283, P = 0.031) and -0.95 (t = 5.842, P = 0.011) respectively.
  
  
  CONCLUSIONS
  miRNA-203 can induce epidermal stem cells to differentiate into sweat gland cells by targeting inhibition of P63 probably.
authors.forename: Zhifang
authors.initials: Z
authors.lastname: Song
authors.forename: Dewu
authors.initials: D
authors.lastname: Liu
authors.forename: Yan
authors.initials: Y
authors.lastname: Peng
authors.forename: Jin
authors.initials: J
authors.lastname: Li
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Zhang
authors.forename: Pu
authors.initials: P
authors.lastname: Ning
authors.forename: Yifeng
authors.initials: Y
authors.lastname: Liu
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D004847:Epithelial Cells; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013545:Sweat Glands / Q000166:cytology*; D014162:Transfection
chemical_list: C546703:MIRN203 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger