Impacts of circACVR2A on Proliferation, Migration, and Epithelial Mesenchymal Transition of Non-Small Cell Lung Cancer Cells by Adjusting miR-486-5p/CCND2 Signaling Axis
WANG Xishi, MI Jie, YANG Chenlu*
The aim of this study is to investigate the effects of circACVR2A (circular RNA activin A receptor 2A type) on the proliferation, migration and EMT (epithelial-mesenchymal transition) of NSCLC (non-small cell lung cancer) cells by regulating miR-486-5p (microRNA-486-5p)/CCND2 (cyclin D2) signaling axis. NSCLC cells HCC827 were cultured in vitro. HCC827 cells were stochastically assigned into Control group, sh-NC group, sh-circACVR2A group, inhibitor-NC group, and miR-486-5p inhibitor group. The qRT-PCR experiment was used to measure the level of circACVR2A, miR-486-5p, and CCND2 mRNA in each group. CCK8 experiment was performed to measure cell proliferation. Cell scratch, Transwell, and flow cytometry experiments were performed to detect cell migration, invasion, and apoptosis. Western blot was performed to measure cell proliferation, apoptosis, migration and invasion, EMT, and CCND2 related protein expression in each group. The dual luciferase assay was used to detect the relationship between circACVR2A and miR-486-5p, as well as miR-486-5p and CCND2. Compared with the Control group and sh-NC group, the sh-circACVR2A group showed decreased circACVR2A, CCND2 mRNA, survival rate, migration rate, invasion number, PCNA, MMP9, Bcl-2, N-cadherin, and CCND2 proteins (P<0.05), and raised miR-486-5p, apoptosis rate, Bax, and E-cadherin (P<0.05). Compared with the sh-circACVR2A group and inhibitor-NC group, the miR-486-5p inhibitor group had no obvious change in circACVR2A, raised CCND2 mRNA, survival rate, migration rate, invasion number, PCNA, MMP9, Bcl- 2, N-cadherin, and CCND2 proteins (P<0.05), and decreased miR-486-5p, apoptosis rate, Bax, and E-cadherin (P<0.05). The dual luciferase assay showed that the mimic NC group had obviously higher luciferase activity in cells than the miR-486-5p mimic group (P<0.05). Downregulation of circACVR2A can inhibit the proliferation, migration, and epithelial mesenchymal transition of NSCLC cells, and promote cell apoptosis, which is associated with miR-486-5p/CCND2 signaling axis.
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