miR-145 modulates IncRNA-ROR and Sox2 expression to maintain human amniotic epithelial stem cell pluripotency and beta islet-like cell differentiation efficiency

摘要

In this study, we observed a great reduction in the expression of the endogenous long noncoding RNA ROR (IncRNA-ROR) and the stem cell transcription factor Sox2, in contrast to a marked increase in miR-145 expression, during the course din 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 IncRNA-ROR and Sox2, silencing their expression. Overexpression of a IncRNA-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 beta islet-like cells. Moreover, the in vitro response of HuAEC-derived beta 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 beta islet-like cell biomarlcers was substantially reduced in HuAECs in the siRNA-ROR transfection group, and their in vivo beta islet-like cell differentiation and insulin release capacities were reduced in a streptozocin-induced diabetic rat model. The experimental results indicate that IncRNA-ROR effectively maintains Sox2 gene expression through competitive binding to miR-145, achieving pluripotency maintenance in HuAECs and regulation of their directed beta islet-like cell differentiation efficiency. (C) 2016 Elsevier B.V. All rights reserved.