title: miR-145 modulates lncRNA-ROR and Sox2 expression to maintain human amniotic epithelial stem cell pluripotency and β islet-like cell differentiation efficiency.
abstract: In this study, we observed a great reduction in the expression of the endogenous long noncoding RNA ROR (lncRNA-ROR) and the stem cell transcription factor Sox2, in contrast to a marked increase in miR-145 expression, during the course of in vitro induced differentiation of human amniotic epithelial stem cells (HuAECs). Bioinformatics analysis and the luciferase reporter assay revealed binding of miR-145 to specific sites in lncRNA-ROR and Sox2, silencing their expression. Overexpression of a lncRNA-ROR-specific siRNA effectively downregulated the expression levels of Sox2 and other stem cell markers in HuAECs while weakening the efficiency of HuAEC differentiation into β islet-like cells. Moreover, the in vitro response of HuAEC-derived β islet-like cells to extracellular stimuli and C-peptide release by these cells were markedly weakened in the siRNA-ROR transfection group. Furthermore, the in vivo expression of β islet-like cell biomarkers was substantially reduced in HuAECs in the siRNA-ROR transfection group, and their in vivo β islet-like cell differentiation and insulin release capacities were reduced in a streptozocin-induced diabetic rat model. The experimental results indicate that lncRNA-ROR effectively maintains Sox2 gene expression through competitive binding to miR-145, achieving pluripotency maintenance in HuAECs and regulation of their directed β islet-like cell differentiation efficiency.
authors.affiliation: Department of Obstetrics, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Zou
authors.affiliation: Department of Obstetrics, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, China; Department of Pathology, Yale University School of Medicine, New Haven, CT 06520, USA. Electronic address: te.liu@yale.edu.
authors.forename: Te
authors.initials: T
authors.lastname: Liu
authors.affiliation: Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Lihe
authors.initials: L
authors.lastname: Guo
authors.affiliation: Laboratoire PROTEE, Bâtiment R, Université du Sud Toulon-Var, 83957, La Garde Cedex, France.
authors.forename: Yongyi
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Neurology, Tongji Hospital, Tongji University School of Medicine, Shanghai 200040, China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Huang
authors.affiliation: Department of Obstetrics, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, China. Electronic address: tduan@yahoo.com.
authors.forename: Tao
authors.initials: T
authors.lastname: Duan
mesh_terms: D000650:Amnion; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002096:C-Peptide / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D003921:Diabetes Mellitus, Experimental / Q000235:genetics / Q000473:pathology / Q000628:therapy; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism; D016381:Islets of Langerhans Transplantation; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D062085:RNA, Long Noncoding / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism*; D013311:Streptozocin
chemical_list: D015415:Biomarkers; D002096:C-Peptide; C000588825:Linc-RNA-RoR, human; C541010:MIRN145 microRNA, human; D035683:MicroRNAs; D062085:RNA, Long Noncoding; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D013311:Streptozocin
keywords: Human amniotic epithelial stem cell; Long noncoding RNA ROR; MicroRNA-145; Sox2; β Islet-like cell