Expression of Recombinant sPDGFR?-Fc in CHO and Its Anti-proliferation Analysis
- VernacularTitle:在CHO细胞中表达重组sPDGFR?-Fc及其抑制细胞增殖的研究
- Author:
Yan WAN
;
Li-Ling LI
;
Qiu-Ling XIE
;
Shu-Jun GUO
;
Li QIN
;
Yong-Cang ZHANG
;
Xiao-Jia CHEN
;
- Publication Type:Journal Article
- Keywords:
sPDGFR? CHO-K1 Anti-proliferation
- From:
China Biotechnology
2006;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Orjective:To obtain recombinant CHO-K1 with expressing sPDGFR? and to identify the biological activities of sPDGFR? secreted in non-serum medium.Methods:Recombinant human sPDGFR? expression vector pIRES-Neo3-sPDGFR?-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 sPDGFR? were analyzed by MTT.Results:sPDGFR?-Fc was cloned into pIRES-Neo3 correctly.The sPDGFR?-Fc expression level in recombinant CHO-K1 cell clones were concordant in between Realtime PCR and Western blot assay.sPDGFR?-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 sPDGFR?-Fc.The sPDGFR?-Fc can inhibit the cell proliferation significantly and it means sPDGFR?-Fc might be a new anti-cancer drug in the future.
