Studies On The Inhibitory Effect Of Mutant Soluble Ectodomain Of FGFR2c(ED-1)on The EGF Signal

Objective Our team successfully constructed mutant soluble ectodomain of human fibroblast growth factor receptor 2 ?c(ED-1)protein.In order to discover what is ED-1's molecular targets,we check if ED-1 was not only able to inhibit FGF signaling by binding to FGF and FGFRs,but also able to inhibit EGF signaling by binding to EGFR,and discuss the possibility that ED-1 may become a double-targeted anti-tumor medicine according to these test results.Methods The expressional levels of FGFRs and EGFR of cancer cell lines were investigated by Real-time Quantitative PCR Detecting System(Q-PCR),in order to primary screening the suitable cell lines.Then,we studied the effect of ED-1 to cancer in vitro and in vivo.In vitro,to explore the role of ED-1 in cancer cells,we focus on the inhibitory effects of ED-1 on cell proliferation and cell cycle.The release levels of FGF-2 and EGF in cancer cell lines were investigated by ELISA.The effects of ED-1 on phosphorylations of FGFRs and EGFR in cancer cells was identified by Western Blot assay.In vivo,ED-1 inhibited the growth of tumors by transplanted tumor model of cancer cell line in Balb/c nude mice.Results According to the Q-PCR results,we chose the breast cancer cell line BT549 and the lung cancer cell line H1299 as the target cell lines.In vitro,the CCK8 results showed that ED-1 inhibited the proliferations of BT549 and H1299.Flow cytometry analysis showed that ED-1 caused BT549 and H1299 cell cycle arrest.Western blot analysis suggested that ED-1 was able to inhibit the expression of p-FGFRs and p-EGFR.In vivo,the transplanted tumors were greatly inhibited by ED-1.Conclutions We successfully constructed mutant soluble ectodomain of human fibroblast growth factor receptor 2 ?c(ED-1)protein,which has the potential to be used as a dual target(FGFRs & EGFR)drug for the treatment of cancers.