Expression of Recombinant sPDGFRα-Fc in CHO and Its Anti-proliferation Analysis

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.