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《Journal of Third Military Medical University》 2012-01
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Promoting effect of RNA-interfered pyruvate dehydrogenase kinase isozyme-1 on 5-fluorouracil-induced apoptosis of colon cancer cells

Xie Ganfeng,Liang Houjie,Tong Jingtao,He Jinxia(Department of Oncology,Southwest Hospital,Third Military Medical University,Chongqing,400038,China)  
Objective To study the effect of pyruvate dehydrogenase kinase isozyme 1(PDK-1) on 5-fluorouracil(5-FU)-induced apoptosis of colon cancer cells.Methods Three interference sequences(interference group) and a negative control sequence of PDK-1 shRNA(control group) were designed to construct pGenesil-1 recombinant plasmid expression vector and pGenesil.1 negative control(pGenesil.1-NC) plasmid vector,respectively.The plasmid was transfected into LS174T cells using liposome and drugs were screened with G418.Effect of RNA interference on 5-FU-induced apoptosis of colon cancer cells was detected by real-time PCR and Western blotting,respectively.Effect of 5-FU on growth and apoptosis of colon cancer cells was detected by MTT assay and flow cytometry,respectively.Results Real-time PCR and Western blotting showed that the 3 interference sequences of PDK-1 shRNA inhibited the expression of PDK-1 in LS174T cells,with a maximum inhibitory rate of over 70%.MTT assay demonstrated that the growth inhibiting rate of LS174T cells interfered with PDK-1 was significantly higher in interference group than in control group 48 h after 5-FU was used(P0.05).Flow cytometry displayed that the apoptosis rate of colon cancer cells was significantly higher in interference group than in control group(P0.05).Conclusion PDK-1 shRNA can down-regulate the expression of PDK-1 in LS174T cells and PDK-1 interference in combination with 5-FU can promote the apoptosis of colon cancer cells.
【Fund】: 国家自然科学基金面上项目(30873015)~~
【CateGory Index】: R735.35
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【Citations】
Chinese Journal Full-text Database 1 Hits
1 Tong Jingtao,Liang Houjie,Chen Jianfang,Jiang Heng,Xie Ganfeng,Liu Ping(Department of Oncology,Southwest Hospital,Third Military Medical University,Chongqing,400038,China);Expression of pyruvate dehydrogenase kinase-Ⅰ in human colon cancer and its significance[J];Acta Academiae Medicinae Militaris Tertiae;2010-03
【Secondary Citations】
Chinese Journal Full-text Database 1 Hits
1 JIAN Ji-yong,ZHANG Yong,ZHU Da-hai(Institute of Basic Medical Science,CAMS & PUMC,Beijing 100005,China);Research progresses on pyruvate dehydrogenase kinases[J];Basic & Clinical Medicine;2008-11
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