Chinese Clinical Oncology ›› 2018, Vol. 23 ›› Issue (1): 13-18.

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Effect of downregulation of miR3917 on proliferation and expression of PER2 in lung cancer A549 cells#br# #br#

  

  1. Department of Respiratory Medicine, Henan Cancer Hospital, the Affiliated Tumor Hospital of Zhengzhou University
  • Received:2017-08-29 Revised:2017-11-17 Online:2018-01-30 Published:2018-06-28

Abstract: ObjectiveTo investigate the effect of microRNA3917 (miR-3917) on the proliferation of lung cancer cells and the expression of biological clock gene Period2 (PER2). 
MethodsThe recovered A549 cells were cultured to logarithmic growth phase. The pEGFPmiR3917siRNA plasmid vector targeting miR3917 was transfected into A549 cells by 10, 20 and 50 nmol/L liposome. The expression of green fluorescent protein was observed under fluorescence microscope after 48 h transfection. According to the experimental design, they were divided into control group, empty load group and interference group. The plasmid concentration in empty load group and interference group was 50 nmol/L. The realtime quantitative PCR (QPCR) was used to detect the relative expression of miR3917 at 48 h after transfection. The mRNA and protein levels of PER2 were detected by QPCR and Western blotting after at 48 h after transfection, respectively. The inhibitive rates of proliferation at 24, 48 and 72 h after transfection were detected by CCK8 assay. Dual luciferase target assay was employed to verify that PER2 was a direct target of miR3917. 
ResultsThe relative expression levels of miR3917 in control group, empty load group and interference group were 1.007±0.018, 1.068±0.084 and 0171±0.052. The relative expression level of miR3917 in the interference group was lower than that in the control group and empty load group, and the difference was statistically significant (P<0.05). The relative expression levels of PER2 were 1.060±0.129, 1.086±0.229 and 2.920±0.927 in the control group, empty load group and interference group, and the protein levels were 0.204±0.046, 0.183±0.043 and 0.512±0.117. The mRNA and protein levels of PER2 in the interference group were higher than those in the control group and empty load group, and the difference was statistically significant (P<0.05). Compared with the control group and empty load group, the proliferative ability decreased in interference group. The inhibitive rates of proliferation were (34.192±5.268)%, (33.527±6.603)% and (30.591±7.788)% at 24, 48, 72 h in interference group, higher than other two groups, and the difference was statistically significant (P<0.05). miR3917 could significantly inhibit the luciferase activity of wildtype PER23’UTR plasmid transfected cells, but had no significant effect on the luciferase activity of mutant PER2-3’UTR plasmid transfected cells. 
ConclusionDownregulation of miR3917 can inhibit the proliferation of lung cancer cells and increase the expression of PER2. miR-3917 has a targeted regulation effect on PER2, and can be used as a new target for the prevention and treatment of lung cancer.

Key words: Lung cancer, A549 cell line, Cell proliferation, MicroRNA3917, Biological clock gene Period2 (PER2)

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