Research Paper Volume 13, Issue 6 pp 8214—8227

Bromodomain-containing protein 4 silencing by microRNA-765 produces anti-ovarian cancer cell activity

miR-765 inhibition upregulates BRD4 and promotes ovarian cancer cell proliferation. Primary ovarian cancer cells (pOC-1 and pOC-2) (A–L) or the established cell lines (CaOV3 and SKOV3) (I–L) were transduced with the lentiviral construct encoding the anti-sense of miR-765 precursor (lv-antagomiR-765) or control anti-sense sequence (lv-antaC), stable cells were established with selection by puromycin. Expression of miR-765 and listed genes was tested by qPCR and Western blotting assays (A, C–E, I, J), with results quantified; the relative BRD4 UTR luciferase reporter activity was tested as well (B). Cell proliferation (by counting EdU-positive nuclei ratio, F, K), cell migration (“Transwell” assays, G, L) and invasion (“Matrigel Transwell” assays, H) were tested, with results quantified. “Pare” stands for the parental control cells. For each assay, n=5 (five replicate well/dishes). Data were presented as mean ± standard deviation (SD). * p

Figure 4. miR-765 inhibition upregulates BRD4 and promotes ovarian cancer cell proliferation. Primary ovarian cancer cells (pOC-1 and pOC-2) (AL) or the established cell lines (CaOV3 and SKOV3) (IL) were transduced with the lentiviral construct encoding the anti-sense of miR-765 precursor (lv-antagomiR-765) or control anti-sense sequence (lv-antaC), stable cells were established with selection by puromycin. Expression of miR-765 and listed genes was tested by qPCR and Western blotting assays (A, CE, I, J), with results quantified; the relative BRD4 UTR luciferase reporter activity was tested as well (B). Cell proliferation (by counting EdU-positive nuclei ratio, F, K), cell migration (“Transwell” assays, G, L) and invasion (“Matrigel Transwell” assays, H) were tested, with results quantified. “Pare” stands for the parental control cells. For each assay, n=5 (five replicate well/dishes). Data were presented as mean ± standard deviation (SD). * p < 0.05 vs. “lv-antaC” cells. Experiments in this figure were repeated five times with similar results obtained.