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中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (07): 1-6    DOI:
    
Expression of recombinant sPDGFRα-Fc in CHO and its anti-proliferation analysis
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Abstract  

Abstract 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.



Received: 24 December 2008      Published: 28 July 2009
ZTFLH:  中图分类号Q786  
Cite this article:

MO Yan, LI Li-Ling, XIE Qiu-Ling, GUO Chu-Jun, QIN Li, ZHANG Yong-Cang, CHEN Xiao-Jia. Expression of recombinant sPDGFRα-Fc in CHO and its anti-proliferation analysis. China Biotechnology, 2009, 29(07): 1-6.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I07/1

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