Studies On Prokaryotic Expression Of Wild-type And Mutant Soluble Ectodomains Of Human Fibroblast Growth Factor Receptor 2IIIc And Their Inhibitory Effects On Tumor

The genes of wild-type and S252W mutant soluble ectodomain of FGFR2â…¢c(sFGFR2â…¢c) were cloned into pET3c,and the recombinant proteins were expressed as inclusion bodies in E.coli.Wild-type and S252W mutant FGFR2â…¢c were refolded by gel filtration chromatography,and the refolded recombinant proteins were purified by heparin affinity chromatography.Both of the purity of target proteins were above 95%.Surface plasmon resonance(SPR) assay was used to examine the affinity of FGF-2 to sFGFR2â…¢c.The results showed that both wild-type and mutant sFGFR2â…¢c could bind FGF-2 specially,and the binding-force of S252W mutant sFGFR2â…¢c and FGF-2 was stronger than wild-type.The inhibitory effects of two kinds of sFGFR2â…¢c on the proliferation of prostate cancer DU145 cells line were examined by MTT assay and Flow Cytometry,The results showed that both sFGFR2â…¢c inhibited evidently the proliferation of DU145 cells.The inhibitory effects of two kinds of sFGFR2â…¢c on growth of tumor were studied by transplanted tumor model of breast cancer cell line MCF-7 in Balb/c nude mice and by lung metastastic model of balb/c breast cancer line 4T1 in balb/c mouse. The results showed that both sFGFR2â…¢c inhibited the growth and metastasis of breast cancer model.Chicken chorioallantoic membrane(CAM) model was used to find that angiogensis were inhibited by two kinds of sFGFR2â…¢c.The inhibitory effect of S252W mutant sFGFR2â…¢c was stronger than wild-type.All above indicated that growth and metastasis of kinds of tumor cell lines can be successfully suppressed by wild-type and S252W mutant sFGFR2â…¢c,and they can be considered as potential broad-spectrum anti-tumor drugs.