A Study On Anti-tumor Activity Of Fusion Protein ATF-EGF-Fc

Objective: The urokinase-type plasminogen activator(urokinase type plasminogen activator,uPA)have been shown to play important roles in tumor growth and angiogenesis. UPAR and Epidermal growth factor receptors (EGFR) are ubiquitous receptors in most malignant tumor cell involved in the processes of tumor proliferation and metastasis. Inhibition of uPA/uPAR and EGFR signaling has demonstrated potent anti-tumor effects. In the present work, we evaluated the activity of ATF-EGFR-Fc, a novel fusion protein of amino -terminal fragment (Amino-Terminal Fragment,ATF) of uPA and EGFR, on inhibition tumor cell in vitro and in vivo.Methods:1 To observe the effects of fusion protein of ATF-EGF-Fc on HUVEC cell proliferation.2 To observe effects of fusion protein of ATF-EGF-Fc on HUVEC cell migration.3 To observe anti-tumor effects of fusion protein of ATF-EGF-Fc through abration assay.4 To observe anti-angiogenic activity of fusion protein of ATF-EGF-Fc through CAM assay. 5 To observe cytotoxic effects of fusion protein ATF-EGF-Fc on tumor cells in vitro.6 To observe anti-tumor effects of fusion protein ATF-EGF-Fc in tumor-bearing nude mice.Results:1 The result of MTT assay showed that the effect of fusion protein on HUVEC proliferation corresponded with the concentration of ATF-EGF-Fc during the range of 100-400nm concentrations, the fusion protein can inhibit the growth of HUVEC cell with a dose-effect depended.2 Both ATF-EGF-Fc and ATF-Fc can reduce HUVEC migration induced by UPA, as compared with control group;.3 Tumor cell proliferation can be inhibited by addting ATF-EGF-Fc to the cell culture plate, as compared with control grouop.4 Addition of ATF-EGF-Fc can greatly inhibit the angiogenesis of CAM as compared with control group.5 MTT assay showed that ATF-EGF-Fc has a toxicity effect on the MCF-7 cell, we can observe that the fusion protein can obviously inhibit the proliferation of MCF-7 cell, which is statistically significant different with control group.6 The inhibitory rate of ATF-EGF-Fc to HepG2 tumor in tumor-bearing nude mice was 85.1%, which is statistically significant different with control group.Conclusions:1 Fusion protein ATF-EGF-Fc can inhibit the proliferation of MCF-7 cell as well as HepG2 cell.2 Through nude mice study, fusion protein ATF-EGF-Fc can strongly inhibit the growth of liver cancer cell, as compared with control group, suggested that it laid a solid foundation to develop a new fusion protein for anti-tumor during clinical practice.