miR-200a inhibits cell proliferation by targeting AP-2γexpression in neuroblastoma cells SK-N-AS / 天津医药

ABSTRACT

Objective To investigate whether miR-200a suppresses cell proliferation by targeting AP-2γexpression, and reveal molecular mechanism that miR-200a functions as a tumor-suppressor in neuroblastoma cells. Methods Dual-luciferase reporter gene assay was employed to examine the effect of miR-200a on AP-2γpromotor luciferase activity. Neu-roblastoma cells were transfected with miR-200a mimics, and the expressions of AP-2γmRNA and protein were detected by RT-PCR and Western blot assay. The effects of AP-2γdown-regulation on cell proliferation were observed after AP-2γshRNA was transfected into neuroblastoma cells. Neuroblastoma cell proliferation was detected by MTS assay after being co-transfected with miR-200a mimics and AP-2γplasmid. Results Results showed that miR-200a could inhibit proliferation of neuroblastoma cells at cell viability (66.33 ± 5.13) compared with that of control group (100 ± 0), and also miR-200a can bind to the 3'untranslated region of AP-2γpromotor and inhibit its luciferase activity with an inhibit ratio at (0.624±0.051). AP-2γmRNA and protein expressions were significantly down-regulated when miR-200a was over-expressed in neuroblas-toma cells. Furthermore, results showed that shRNA-mediated down-regulation of AP-2γthat suppressed the cell prolifera-tion of neuroblastoma at (62.5±2.4) by comparing with the control group (100±0). Moreover, restoring AP-2γexpression re-versed the effect of miR-200a with a cell viability suppression at (92.4±1.4). Conclusion miR-200a suppresses cell prolif-eration by targeting AP-2γexpression in neuroblastoma cells.