Ectopic expression of miR-181a suppresses growth and induces apoptosis of REH cells.

REH cells were infected with lentiviral vector expressing the negative control shRNA (NC) or miR-181a (181A1). (A) Relative miR-181a levels were determined by qRT-PCR. (B) Growth curve was determined as following: Cells were cultured for 48, 96, and 144 hours and then assessed with the MTT assay. (C) Cells were seeded at a density of 1 × 105 cells/mL and cultured. After 72 hours, cells were stained with trypan blue, and viable cells were counted. (D) Apoptosis was assessed by flow cytometric analysis of annexin V/propidium iodide (PI) staining of lentivirus-transduced cells (left). Representative histograms demonstrate the proportion of annexin V-positive cells (right). (E) Biparametric BrdU/DNA analysis: During the last 30 minutes of culture, 1 mM BrdU was added to the cells, and then the cells were stained with anti-BrdU and 7-aminoactinomycin D (7-AAD) and detected by flow cytometry (left). The percentage of cells in each of the cell-cycle phases G0/G1, S, and G2+M was quantified (right). Bars show the mean ± SD from three independent experiments. 181A1 vs. NC *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 (ANOVA); NC vs. REH ##P ≤ 0.01.