LI Ying,HE Wei,ZHANG Bin,WU Jun,WANG Ru-peng,LIN Zi-hua.Construction of human papilloma virus 16 E7 eukaryotic expression vector and its effect on proliferation and invasion of A431 cells[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(09):818-821.
LI Ying,HE Wei,ZHANG Bin,WU Jun,WANG Ru-peng,LIN Zi-hua.Construction of human papilloma virus 16 E7 eukaryotic expression vector and its effect on proliferation and invasion of A431 cells[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(09):818-821.DOI:
Objective To construct an eukaryotic expression vector encoding human papilloma virus E7
and investigate the effect of E7 overexpression on the proliferation and invasion of human epithelial carcinoma cell line A431. Methods After total RNA from Caski cells was obtained by Tripurer
cDNA was amplified by RT-PCR and then cloned into pEGFP-N1 vector using gene engineering technology. Recombinant expression vector pEGFP-E7 was confirmed by restriction digestion and sequencing
and then tran
sfected into A431 cells by liposome. Expression of E7 protein was detected by Western blotting. WST-1 assay and Transwell system were used to observe the growth and invasion of A431 cells. Cell cycle was measured by flow cytometry
and the expression of TGF-β in the supernatant was measured by ELISA. Results The recombinant expression vector pEGFP-E7 was successfully constructed. Western blotting indicated that E7 protein was overexpressed in A431 cells. Overexpression of E7 significantly increased cell proliferation and invasion. Flow cytometry showed there were more cells arresting in S phase and lesser in G
1
phase. Overexpression of E7 significantly improved the secretion of TGF-β in A431 cells. Conclusion Overexpression of E7 enhances tumor cell proliferation and invasion
although TGF-β secretion is increased at the same time.