Study on effect of miR-107 targeting CCNE1 on function of human non-small cell lung cancer cell line A549 / 重庆医学

ABSTRACT

Objective To investigate the effect of miR-107 on the function of human non-small cell lung cancer cell line A549 and its possible target genes.Methods The experiment was divided into the liposome+miR-107 mimics overexpression group(OV-miR-107 group),liposome+miR-107 inhibiting mimics downregulation group(KD-miR-107 group)and liposome+ negative control mimics group(NC group).The cell transfections of siRNA were siRNA-1,siRNA-2 and siRNA-3 respectively.The cellular prolifer-ation capacity was detected by MTT assay.The cellular cycle was detected by flow cytometry.The dual luciferase reporter gene as-say was performed to detect the downstream target gene of miR-107.The real time quantitative reverse transcription PCR and Western blot were respectively employed to examine mRNA and protein expression levels of downstream target gene.Results miR-107 inhibited the proliferation of A549 cells in a time-and dose-dependent manner(P<0.05).miR-107 arrested more A549 cells at the G0G1phase,and the proportions of S phase and G2M phase were decreased(P<0.05).miR-107 combinded with the 246 bp-253 bp of CCNE1 3′-untranslated region,and decreased mRNA and protein expression of CCNE1(P<0.05);after down-regulating CC-NE1 in A549 cells,siRNA-2 inhibited cellular proliferation(P<0.05)and blocked the cells to stay at G0G1phase(P<0.05).Con-clusion miR-107 inhibits the proliferation of human non-small cell lung cancer cell line A549 and regulates the cellular cycle by tar-geting CCNE1.