FGFR2IIIc胞外段的原核表达及其对前列腺癌细胞增殖的抑制作用

将FGFR2IIIc胞外段基因转入pET3c载体，并通过大肠杆菌使其以包涵体形式表达。经透析法复性和肝素亲和层析纯化，得到纯度95%以上的目的蛋白。通过免疫共沉淀方法证实复性后的FGFR2IIIc胞外段与FGF2之间可特异性结合，并对前列腺癌DU145细胞的增殖具有明显的抑制作用（MTT法）。免疫印迹结果显示FGFR2IIIc胞外段可显著下调DU145细胞膜上FGFRs的磷酸化水平，提示由于有效阻断FGFs的细胞内信号通路，从而导致FGFR2IIIc胞外段对肿瘤细胞增殖的抑制。

Prokaryotic Expression of Soluble Ectodomain of Human Fibroblast Growth Factor Receptor 2IIIc and Its Inhibitory Effect on the Proliferation of DU145 Cells

Abstract The gene of soluble ectodomain of FGFR2IIIc (sFGFR2IIIc) was cloned into pET3c, and expressed as inclusion bodies in E.coli. The inclusion bodies were refolded by dialysis and then purified. The purity of sFGFR2IIIc was more than 95% after heparin affinity chromatography. The result of co-IP indicated that the product was able to bind FGF2, and the proliferation of prostate cancer DU145 cells line was inhibited by sFGFR2IIIc in a time-dependent and dose-dependent manner by MTT assay. Moreover, the phosphorylation of FGFRs on the cell membrane could be downregulated dramatically after treatment with sFGFR2IIIc. The results suggested that sFGFR2IIIc could inhibit the proliferation of DU145 cells through blocking the cell signaling of FGFs.