LncRNA PSMA3-AS1 mediates proliferation,apoptosis and epithelial mesenchymal transition of lung cancer cells by regulating the miR-140-3p/DDX5 axis
Objective To explore the regulatory effects of long non-coding RNA proteasome subunit alpha type-3 antisense RNA 1(PSMA3-AS1)on lung cancer cell proliferation,apoptosis,and epithelial mesenchymal transition(EMT)by regulating the miR-140-3p/DEAD box p68 RNA helicase(DDX5)axis.Methods Quantitative reverse transcription polymerase chain reaction(qRT-PCR),Western blot,and immunohistochemistry were applied to detect the expressions of PSMA3-AS1,miR-140-3p,and DDX5 in 40 lung cancer tissues and lung cancer cell lines.A549 cells were treated with blank control,or transfected with si-NC,si-PSMA3-AS1,si-PSMA3-AS1+inhibitor-NC,or si-PSMA3-AS1+miR-140-3p inhibitor.Transfection efficacy was tested by qRT-PCR.Cell proliferation and apoptosis were detected by MTT assay and flow cytometry,respectively.Cell migration and invasion were detected by Transwell assay.Protein expressions of PCNA(proliferating cell nuclear antigen),Bax(Bcl-2 associated X protein),MMP2(matrix metalloproteinase),Vimentin,E-cadherin and DDX5(DEAD box protein 5)were examined by Western blot.The targeting relationship between miR-140-3p and PSMA3-AS1,and that between miR-140-3p and DDX5 was verified by dual-luciferase reporter assay.A lung cancer xenograft model was created in nude mice implanted with lung cancer cells transfected with si-NC or si-PSMA3-AS1.Tumor weight and volume were measured.Relative level of miR-140-3p in mouse lung cancer tissues was detected by qRT-PCR.Protein expressions of Ki-67 and DDX5 in mouse lung cancer tissues were examined by immunohistochemical staining.Results Upregulated mRNA level of PSMA3-AS1,upregulated protein level of DDX5 and downregulated mRNA level of miR-140-3p were detected in lung cancer tissues and cell lines(P＜0.05).Knockdown of PSMA3-AS1 significantly inhibited the proliferation,migration and invasion of A549 cells,promoted apoptosis,downregulated PCNA,MMP-2,Vimentin and DDX,and upregulated miR-140-3p,Bax and E-cadherin(P＜0.05).Knockdown of miR-140-3p weakened the role of silenced PSMA3-AS1 in inhibiting the proliferation,migration and invasion of A549 cells and promoting apoptosis(P＜0.05).Dual-luciferase reporter assay confirmed the targeting relationship between miR-140-3p and PSMA3-AS1,and that between miR-140-3p and DDX5(P＜0.05).In vivo data revealed that knockdown of PSMA3-AS1 significantly reduced tumor weight and volume in nude mice,downregulated Ki-67 and DDX5,and upregulated miR-140-3p(P＜0.05).Conclusion Knock-down of PSMA3-AS inhibits the proliferation and EMT of lung cancer cells,and promotes cell apoptosis by regulating the miR-140-3p/DDX5 axis.
long non-coding RNA proteasome subunit alpha type-3 antisense RNA 1(lncRNAs PSMA3-AS1)MiR-140-3p/DEAD-box protein 5(DDX5)axislung cancerproliferationapoptosisepithelial mesenchymal transition
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