Chinese Clinical Oncology

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Effects of up-regulated expression of miR-125a on cell apoptosis of human lung cancer A549 cells and possible mechanisms

HU Jinhua,WU Yuquan,CHEN Qingyong,ZHAO Yuanyuan,JIAO Demin.   

  1. epartment of Internal Medicine,the 117th Hospital of PLA,Hangzhou 310013,China
  • Received:2012-11-08 Revised:2012-12-25 Online:2013-03-31 Published:2013-03-31

Abstract: bjective To explore the effect of up-regulated expression of microRNA-125a(miR-125a) on cell apoptosis of human lung cancer A549 cells and possible mechanisms. Methods The miR-125a sequence was synthesized and cloned into eukaryotic expression plasmid pGenesil-1 to construct recombinant plasmid pGenesil-miR-125a.Meanwhile,a control plasmid pGenesil control was constructed. The above plasmids were transfected into human lung cancer A549 cells using liposomes and divided into three groups: A549 cells transfected with pGenesilmiR-125(A549-miR-125a group),A549 cells transfected with pGenesil control(A549-control group)and A549 cells without transfection(A549). Quantitative realtime polymerase chain reaction(QPCR) was used to measure the levels of miR-125a(normalized to U6 mRNA levels) after transfection. The percentage of apoptosis was determined by Annexin V-FITC/PI double staining with flow cytometry analysis. Western blot was employed to analyze the protein levels of p53. Results The recombinant plasmid pGenesil-miR-125a was verified by restriction analysis and DNA sequencing. The level of miR-125a in A549-miR-125a group was 2.72±0.41,higher than 0.97±0.16 of A549-control group and 0.96±0.11 of A549 group(P<0.01). The apoptotic rate of A549-miR-125a group was(31.04±2.48)%,higher than (6.91±0.72)% of A549-control group and (6.73±0.56)% of A549 group(P<0.05). The level of p53 protein in A549-miR-125a group was 3.91±0.46,higher than 1.01±0.06 of A549 control group and 0.99±0.04 of A549 group(P<0.01). Conclusion Up-regulated expression of miR-125a can promote the apoptosis of lung cancer A549 cells with the possible mechanism of the up regulation of p53.

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