miR-30a-5p Targeted E2F7 to Block A549 Cell Cycle and Inhibit Its Proliferation
Objective To investigate the mechanism of miR-30a-5p targeting E2F7 in regulating cell cycle and proliferation of non-small cell lung cancer(NSCLC).Methods The relative expression levels of miR-30a-5p in non-small cell lung cancer A549 cells and human normal lung epithelial cells BEAS-2B were verified by quantitative real-time PCR(qRT-PCR).The target gene and binding site of miR-30a-5p were predicted by online software,and the targeting relationship between miR-30a-5p and E2F7 mRNA 3'UTR region was further verified by dual luciferase reporter gene assay.Secondly,A549 cells were transfected with miR-30a-5p mimics and miR-30a-5p inhibitor,and the proliferation of A549 cells was detected by CCK-8 cell counting kit.The apoptosis and cell cycle of A549 cells were detected by flow cytometry.qRT-PCR and Western blot were used to detect the mRNA and protein expression levels of cell cyclin-related genes and lung adenocarcinoma-related genes.Results The expression level of miR-30a-5p in A549 cells was significantly lower than that in BEAS-2B cells,and miR-30a-5p inhibited the expression of E2F7 by targeting the 514-520 site of E2F7 mRNA 3'UTR.Transfection of miR-30a-5pmimics significantly inhibited the proliferation and promoted the apoptosis of A549 cells.A549 cell cycle was arrested at G1phase.It also significantly regulated the mRNA and protein expression levels of lung adenocarcinoma related genes TFDP3,CDK1,E2F8,CCNA2 and CDC6.Conclusion miR-30a-5p inhibits the proliferation and cell cycle of non-small cell lung cancer A549 cells by negatively regulating the expression of E2F7 and the mRNA and protein expressions of TFDP3,CDK1,E2F8,CCNA2 and CDC6.
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