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《Basic & Clinical Medicine》 2008-01
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Cloning of mouse GRIM-19 gene and its apoptotic effect on prostate cancer rm-1 cells

SHAO Yue-ting1,ZHANG Ling1,JI Kun1,LIU Ya-nan1,LI Yang1,HU Jia-di3,ZHAO Xue-jian2(1.Department of Pathophysiology of Basic Medicine College of Jilin University;2.Prostate Diseases Prevention and Treatment Researoh Certer,Jilin University,Changchun 130021,China;3.University of Maryland School of Medicine,MD 21201,USA)  
Objective To construct pcDNA3.1-GRIM-19 eukaryotic expression vector and to investigate its function in mouse prostate cancer rm-1 cells.Methods RT-PCR was performed on total RNA extracted from mouse spleen tissue to obtain the cDNA of GRIM-19,which was inserted into pMD-18T vector.DNA sequencing was tested before the amplified products were cloned into pcDNA3.1.The recombinant vector was transfected into rm-1 cells and its expression was examined by RT-PCR.The apoptotic effects of cells were observed by laser scanning confocal microscope(LSCM)and DNA ladder assay.Then,using the RT-PCR detected caspase-3 activity in rm-1 cells after transfection.Results The amplified products were confirmed as the cDNA of rm-1 by DNA sequencing,being capable of expression in rm-1 cells.Typical apoptosis was observed by LSCM and DNA ladder in recombinant plasmid group after 48h of transfection in rm-1 cells.Overexpression of GRIM-19 increased caspase-3 expression.Conclusion The eukaryotic expression vector for GRIM-19 was successfully constructed,which can be expressed in rm-1 cells and can induce the cell apoptosis.Its apoptosis mechanism may related to caspase-3 activity.
【Fund】: 国家科技部国际科技合作重点项目(2004DFB02000);; 中日政府间专项技术合作(第59号)
【CateGory Index】: R737.25
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