[Effect of over expression of HPV18 E2 protein on apoptosis and secretion of macrophages].

AIM: To study the effect of the over expression of GFP-E2, GFP-TAD(N-extremity domain of HPV18 E2) and GFP-DBD (C-extremity domain of HPV18 E2) on the apoptosis and secretion of macrophages and to further explore the contribution of E2 gene to the uterine cervix cancer. METHODS: TAD or DBD gene was amplified from pEGFP-C1/HPV18 E2 by PCR respectively and then cloned into pEGFP-C1 vector. After the transfection of recombinant plasmids or pEGFP-C1 into the macrophages, their expression was examined by fluorescent microscopy and Western blot. The cytokine content of TNF-alpha or IL-1alpha in the culture medium was tested quantitatively with ELISA kit respectively. The stained macrophages were observed and their apoptosis rate was tested by flow cytometry. RESULTS: After transfected into macrophages, GFP-E2 fusion protein was mainly located in cytoplasma while GFP-DBD fusion protein was completely located in nuclei and GFP-TAD fusion protein was completely located in cytoplasma. The overexpression of GFP-E2 or GFP-TAD increased the level of TNF-alpha and IL-1alpha and upregulate the apoptosis rate of macrophages. Furthermore, the effect of GFP-TAD was obvious except on IL-1beta level but the overexpression of GFP-DBD did not show the same effect. CONCLUSION: The overexpression of GFP-E2 or GFP-TAD fusion protein can induce the apoptosis macrophages and upregulate TNF-alpha or IL-1beta secretion of macrophages.