Journal: PLoS ONE

Article Title: Let-7b Inhibits Human Cancer Phenotype by Targeting Cytochrome P450 Epoxygenase 2J2

doi: 10.1371/journal.pone.0039197

Figure Lengend Snippet: A , proliferation assay via Cell-Light™ EdU DNA Cell Proliferation Kit showing decreased proliferation rate of MDA-MB-435 and SK-MES-1 cells treated with let-7b compared to the cells treated with random let-7b or control. The blue-stained cells were stained by Hoechst, and the red were EdU add-in cells. EdU-positive cells were calculated as (EdU add-in cells/Hoechst-stained cells)×100%. Columns, mean of three experiments; bars, SD. *, P <0.05. B , percentage of apoptotic cells was increased in MDA-MB-435 and SK-MES-1 cells treated with let-7b. Apoptosis measured by annexin V-FITC (x-axis) and propidium iodide staining (y-axis). Percentages of apoptotic cells (percentage of cells in the upper-right quadrant (annexin V-positive, PI-negative) plus cells in the low-right quadrant (annexin V-positive, PI-positive) in total cell number) are given under the relevant graph. Columns, mean of three experiments; bars, SD. *, P <0.05. C–D , percentages of EdU-positive cells and apoptotic cells were analyzed in HeLa and Tca-8113 cells transfected with let-7b or random let-7b. E–F , MDA-MB-435 and SK-MES-1 cells transfected with let-7b were treated with C26 (specific CYP2J2 inhibitor, 10 µM) or 14,15-EET (250 nM). 48 h later, percentages of EdU-positive cells and apoptotic cells were analyzed. Columns, mean of three experiments; bars, SD. *, P <0.05. G , let-7b overexpression influenced expression of tumor-related genes in MDA-MB-35 cells. Let-7b overexpression in MDA-MB-435 cells significantly downregulated PI3K, pAkt, and pERK but increased Bax and nm-23 expression. The data shown were repeated three times.

Article Snippet: Cell proliferation was monitored using Cell-Light EDU DNA Cell Proliferation Kit (Ribobio, China).

Techniques: Proliferation Assay, Control, Staining, Transfection, Over Expression, Expressing