Effects circ-CPA4 on the Proliferation,Apoptosis,Migration and Invasion of Non-Small Cell Lung Cancer Cells by Regulating the miR-140-3p/SGK1 Axis
Objective:To investigate the effects of circular RNA carboxypeptidase A4(circ-CPA4)on proliferation,apoptosis,mi-gration and invasion of non-small cell lung cancer(NSCLC)cells by regulating miR-140-3p/serum and glucocorticoid-induced protein kinase-1(SGK1)axis.Methods:H1299 cells were grouped into five groups:Ct group,si-NC group,si-circ-CPA4 group,si-circ-CPA4 combined with inhibitor-NC group and si-circ-CPA4 combined with miR-140-3p inhibitor group.The real time fluorescence quantitative reverse transcription polymerase chain reaction(qRT-PCR)was performed to detect the expressions of circ-CPA4 and miR-140-3p.The flow cytometry was performed to detect cell apoptosis.The cell counting Kit(CCK-8)assay was performed to detect cell proliferation.Migration was detected by scratch-healing experiment.Invasion was detected by Transwell.The western blot was performed to detect the expressions of SGK1,cysteine protease 3(Caspase-3),cyclin D1(CyclinD1),and matrix metalloproteinase 2(MMP-2)proteins.The tar-geting relationship between circ-CPA4 and miR-140-3p,miR-140-3p and SGK1 were detected by dual-luciferase reporter gene assay.The cell suspensions of the above five groups were subcutaneously injected into nude mice respectively,and after 30 days,the nude mice were sacrificed,the tumors were separated,and the tumor mass was weighed.Results:In NSCLC tissues and cells,circ-CPA4 and SGK1 proteins were highly expressed,and miR-140-3p was lowly expressed.Compared with Ct group and si-NC group,the circ-CPA4,scratch healing rate,OD450 value,number of invasive cells,SGK1,CyclinDl,MMP-2 protein expressions and tumor mass in nude mice were sig-nificantly decreased in si-circ-CPA4 group,the expression of miR-140-3p,the rate of apoptosis,and the protein expression of Caspase-3 were increased significantly(P＜0.05).Compared with the si-circ-CPA4 combined with inhibitor-NC group,the change trend of the corre-sponding indicators in the si-circ-CPA4 combined with miR-140-3p inhibitor group was opposite to the above(P＜0.05).The silencing circ-CPA4 could target down-regulate miR-140-3p expression,down-regulation of miR-140-3p can target and promote SGK1 protein expression.Conclusion:Silencing circ-CPA4 inhibits SGK1 protein expression by up-regulating miR-140-3p,thereby inhibiting prolifer-ation,migration and invasion of NSCLC cells,and promoting cell apoptosis.
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