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


CONCLUSIONS
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.
Peng
Jun
J
Institute of Pathogenic Biology, University of South China, Hengyang 421001, China. junpeng7758@yahoo.com.cn
Zhu
Cui-ming
CM
Yu
Min-jun
MJ
Cao
Qing-xiang
QX
Wang
Xin
X
Wan
Yan-ping
YP
D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D052161:Human papillomavirus 18 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D053583:Interleukin-1beta; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D049452:Green Fluorescent Proteins