MiRNAs inhibiting lung adenocarcinoma cell proliferation by targeting oncogene TRIB2

【作者】 谢书阳； 李有杰； 张超； 王萍玉； 辛佳璇； 李新新；

【Author】 Key Laboratory of Tumour Molecular Biology in Binzhou Medical University.Department of Biochemistry and Molecular Biology,Binzhou Medical University;

【机构】 Key Laboratory of Tumour Molecular Biology in Binzhou Medical University.Department of Biochemistry and Molecular Biology,Binzhou Medical University；

【摘要】 miRNAs are small noncoding RNAs that play important roles in several human diseases,including cancer.To explore the actions of miR-511 and miR-1297 on A549 cells,their targeting sites of trib2 gene were predicted by microRNA analysis software,and pcDNA-GFP-TRIB2-3’UTR vector was constructed to investigate the interaction between TRIB2 and miR-511/1297.The GFP expression was estimated by fluorescence microscopy and flow cytometry after A549 cells were transfected with miR-511(or miR-1297) and pcDNA-GFP-TRIB2-3’UTR vector.The expression of GFP in the miR-511 and miR-1297 groups was significantly downregulated contrasted with the NC control group.The expression of TRIB2 was further detected to be decreased after miR-511(or miR-1297) treatment by western blotting.After miR-511 and miR-1297 treatment,A549 cell proliferation inhibition rate was higher using MTT test,and the apoptotic rate was increased with Annexin V-FITC/PI dual staining analysis compared to NC treatment group.As the downstream factor of TRIB2,C/EBPa expression was enhanced after miR-511(or miR-1297) treatment.Our results illustrated that miR-511 and miR-1297 act as tumour suppressor genes,which could induce A549 cell apoptosis by targeting TRIB2 and further increasing C/EBPa expression.更多还原

【Abstract】 miRNAs are small noncoding RNAs that play important roles in several human diseases,including cancer.To explore the actions of miR-511 and miR-1297 on A549 cells,their targeting sites of trib2 gene were predicted by microRNA analysis software,and pcDNA-GFP-TRIB2-3’UTR vector was constructed to investigate the interaction between TRIB2 and miR-511/1297.The GFP expression was estimated by fluorescence microscopy and flow cytometry after A549 cells were transfected with miR-511(or miR-1297) and pcDNA-GFP-TRIB2-3’UTR vector.The expression of GFP in the miR-511 and miR-1297 groups was significantly downregulated contrasted with the NC control group.The expression of TRIB2 was further detected to be decreased after miR-511(or miR-1297) treatment by western blotting.After miR-511 and miR-1297 treatment,A549 cell proliferation inhibition rate was higher using MTT test,and the apoptotic rate was increased with Annexin V-FITC/PI dual staining analysis compared to NC treatment group.As the downstream factor of TRIB2,C/EBPa expression was enhanced after miR-511(or miR-1297) treatment.Our results illustrated that miR-511 and miR-1297 act as tumour suppressor genes,which could induce A549 cell apoptosis by targeting TRIB2 and further increasing C/EBPa expression.更多还原