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miR-25 promotes cell proliferation by targeting RECK in human cervical carcinoma He La cells

QIU Gang;FANG Baoshuan;XIN Guohong;WEI Qiang;YUAN Xiaoye;WU Dayong;Second Department of Oncology,Hebei General Hospital;Third Department of Oncology,Hebei General Hospital;Depatment of Nuclear Medicine,Hebei General Hospital;Second Department of Geriatrics,Hebei General Hospital;  
Objective To investigate the effect of miR-25 on the proliferation of human cervical carcinoma He La cells and its association with reversion-inducing cysteine-rich protein with Kazal motifs(RECK).Methods The recombinant plasmids of pc DNATM6.2-GW-pre-miR-25,pmir GLO-RECK-WT,pmir GLO-RECK-MT and anti-miR-25 were constructed,and their transfection efficiencies into He La cells were identified by real-time quantitative PCR(qRT-PCR).The potential proliferationstimulating function of miR-25 was analyzed by MTT assay in He La cells.Furthermore,the target effect of miR-25 on the RECK was determined by dual-luciferase reporter assay system,qRT-PCR and Western blotting.Results Sequence analysis demonstrated that the recombinant plasmids of pc DNATM6.2-GW-pre-miR-25 and pmir GLO-RECK-WT,pmir GLO-RECK-MT were successfully constructed,and qRT-PCR revealed that the transfection efficiencies of pre-miR-25 and anti-miR-25 were desirable in He La cells.MTT assay showed that miR-25 over-expression promoted the proliferation of He La cells.In addition,the luciferase activity was significantly reduced in He La cells cotransfected with pre-miR-25 and RECK-WT.The qRT-PCR and Western blotting indicated that the expression level of RECK was up-regulated in He La cells transfected with anti-miR-25 at the transcriptional and posttranscriptional levels.Conclusion miR-25 could promote cell proliferation by targeting RECK in He La cells.
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