OBJECTIVETo investigate miR-20a expression in human glioma and normal brain tissues and its effect on the proliferation of glioma cells in vitro.

METHODSThe expression of miR-20a was detected in human normal brain tissues and glioma tissues by real-time RT-PCR. miR-20a mimics were synthesized and transfected into U251 cells via liposome, and the cell proliferation were detected using MTT assay and flow cytometry.

RESULTSThe glioma tissues showed significantly up-regulated expression of miR-20a compared with normal brain tissues (P=0.035). The expression level of miR-20a was higher in high-grade than in low-grade gliomas. miR-20a mimics significantly enhanced the proliferation of U251 cells and the percentage of S-phase cells.

CONCLUSIONmiR-20a shows potent effect in promoting the growth of glioma cells, suggesting its important role in the pathogenesis of human glioma.

Adult ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Male ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Real-Time Polymerase Chain Reaction ; Young Adult

Background and Objectives: The search for a suitable alternative for urethral defect is a challenge in the field of urethral tissue engineering. Induced pluripotent stem cells (iPSCs) possess multipotential for differentiation. The in vitro derivation of urothelial cells from mouse-iPSCs (miPSCs) has thus far not been reported. The purpose of this study was to establish an efficient and robust differentiation protocol for the differentiation of miPSCs into urothelial cells. Methods: and Results: Our protocol made the visualization of differentiation processes of a 2-step approach possible. We firstly induced miPSCs into posterior definitive endoderm (DE) with glycogen synthase kinase-3β (GSK3β) inhibitor and Activin A. We investigated the optimal conditions for DE differentiation with GSK3β inhibitor treatment by varying the treatment time and concentration. Differentiation into urothelial cells, was directed with all-trans retinoic acid (ATRA) and recombinant mouse fibroblast growth factor-10 (FGF-10). Specific markers expressed at each stage of differentiation were validated by flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR) assay, immunofluorescence staining, and western blotting Assay. The miPSC-derived urothelial cells were successfully in expressed urothelial cell marker genes, proteins, and normal microscopic architecture. Conclusions We built a model of directed differentiation of miPSCs into urothelial cells, which may provide the evi-dence for a regenerative potential of miPSCs in preclinical animal studies.