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《China Biotechnology》 2009-07
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Expression of Recombinant sPDGFRα-Fc in CHO and Its Anti-proliferation Analysis

WAN Yan LI Li-ling XIE Qiu-ling GUO Shu-jun QIN Li ZHANG Yong-cang CHEN Xiao-jia(National Engineering Research Center of Genetic Medicine,Guangdong Provincial Key Laboratory of Bioengineering Medicine,Bioengineering Institute of Jinan University,Guangzhou 510632,China)  
Orjective:To obtain recombinant CHO-K1 with expressing sPDGFRα and to identify the biological activities of sPDGFRα secreted in non-serum medium.Methods:Recombinant human sPDGFRα expression vector pIRES-Neo3-sPDGFRα-Fc was constructed and then transfected into CHO-K1 cells by using LipofectamineTM 2000.After screened with G418 in 8 weeks,some monoclone cells were selected randomly to amplify in 96-well-plate to 24-well-plates,and then to identify positive cell clones by RT-PCR.Furthermore,the candidate cell clones were test by Real-Time PCR and Western blot assays.Finally,anti-proliferation activities of the expressed sPDGFRα were analyzed by MTT.Results:sPDGFRα-Fc was cloned into pIRES-Neo3 correctly.The sPDGFRα-Fc expression level in recombinant CHO-K1 cell clones were concordant in between Realtime PCR and Western blot assay.sPDGFRα-Fc obtained from cultured non-serum medium of positive CHO-K1 could significantly inhibit proliferation of vascular endothelial cell.Conclusion:Successed to select recombinant CHO-K1 cell lines with high expressed sPDGFRα-Fc.The sPDGFRα-Fc can inhibit the cell proliferation significantly and it means sPDGFRα-Fc might be a new anti-cancer drug in the future.
【Fund】: 国家“863”计划重大专项子课题(2002AA22344E);; 广东省科技攻关项目(200813030301349)资助项目
【CateGory Index】: Q786
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【Citations】
Chinese Journal Full-text Database 2 Hits
1 ZHANG Guo-qiang, LIU Zhi-gang, YU Wei-yuanInstitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China;Latest Developments in Highly Efficient Expression Vectors in Mammalian Cells[J];Letters in Biotechnology;2005-01
2 ZHANG Xiu-hua LIN Li-ping DING Jian (Division of Anti-tumor Pharmacology,State Key Laboratory of Drug Research,Shanghai Institute of Materia Medica,Shanghai Institutes for Biological Science,Chinese Academy of Sciences,SHANGHAI 201203,China);Clinical development of tyrosine kinase inhibitors targeted platelet-derived growth factor receptor[J];Chinese Journal of New Drugs and Clinical Remedies;2006-06
【Co-citations】
Chinese Journal Full-text Database 3 Hits
1 WAN Yan,LI Li-ling,CHEN Xiao-jia(Bioengineering Institute of Jinan University,Guangzhou 510632,China);A new method to tetermine the copy numbers of exogenous gene by absolute quantitative Real-Time PCR assay[J];Journal of Jinan University(Natural Science & Medicine Edition);2009-03
2 XIE Hao~*,LI Qi-Chang,GUO Xiao-Ming(Department of Biological Science and Biotechnology,Institute of Science,Wuhan University of Technology,Wuhan 430070,China);Overexpression and Purification of Membrane Transport Protein with Native Functions[J];Chinese Journal of Biochemistry and Molecular Biology;2007-12
3 CHE Wen-jun, LI Zhi-yu, YOU Qi-dong(Department of Medicinal Chemistry, China Pharmaceutical University, Nanjing 210009, China);Current Targets for Anticancer Drug Discovery and the Related Drugs[J];Progress in Pharmaceutical Sciences;2007-06
【Secondary Citations】
Chinese Journal Full-text Database 2 Hits
1 LIU Guo Qi,WANG Hai Tao(Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China).;Optimized Strategies to Hyperexpress Recombinant Protein in Chinese Hamster Ovary Cells.[J];PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS;2000-05
2 LAI Da-Zhi QI Lian-Quan YU Chang-Ming WANG Hai-Tao CHEN Wei * (Beijing Institute of Microbiology and Epidemiology, Beijing\ 100071, China);Modification of Chinese Hamster Ovary Cells[J];Chinese Journal of Biotechnology;2002-04
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