Abstract
Objective
To study the interaction of human papillomavirus type 16 (HPV16 E6) protein and human death domain associated protein (hDaxx) and its effect on apoptosis of HeLa cells to provide the experimental basis for exploring the oncogenic mechanism of HPV16 E6 protein.
Methods
Recombinant vector of pGADT7/E6 or pGBKT7/hDaxx was constructed. The interaction of E6 protein and hDaxx was detected by yeast two-hybrid system. Their expression in yeast was detected by Western blotting. The eukaryotic plasmids of E6 and hDaxx were co-transfected into HeLa cells. Apoptosis was induced by 5-FU. The apoptotic rate was measured by flow cytometry (FCM).
Results
E6 protein had intracellular interaction with hDaxx. The apoptotic rate was rising with the increase in the transfection quantity of pcDNA3.1 (-) / hDaxx in pcDNA3.1 (-) /E6 and pcDNA3.1 (-) / hDaxx co-transfected cells. The difference was significant (P < 0. 01).
Conclusion
There is intracellular interaction between HPV16 E6 protein and hDaxx. The over-expression of hDaxx can increase the sensitivity of E6 protein positive HeLa cells to 5-FU. The effect was in a dose dependent manner. HPV16 E6 protein inhibited the apoptosis of HeLa cells by interacting with hDaxx.
Similar content being viewed by others
References
Pretet JL, Charlot JF, Mougin C. Virological and carcinogenic aspects of HPV. Bull Acad Natl Med, 2007, 191: 611–623.
Hiller T, Stubenrauch F, Iftner T. Isolation and functional analysis of five HPVE6 variants with respect to p53 degradation. J Med Virol, 2008, 80: 478–483.
Gurer C, Berthoux L, Luban J. Covalent modification of human immunodeficiency virus type 1 p6 by SUMO-1. J Virol, 2005, 79: 910–917.
Cantrell SR, Bresnahan WA. Human cytomegalovirus (HCMV) UL82 gene product (pp71) relieves hDaxx-mediated repression of HCMV replication. J Virol, 2006, 80: 6188–6191.
Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics, 2002. CA Cancer J Clin, 2005, 55: 74–108.
Stanley MA, Pettm R, Coleman N. HPV: from infection to cancer. B iochem Soc Trans, 2007, 35: 1456–1460.
Jiang YL, Wang J, Yu MJ, et al. Construction of Human Papillomavirus type 16 E6 eukaryotic expression vector and its expression in eukaryotic cells. J Nanhua Univ (Medical edition) (Chinese). 2006, 34: 505–508.
Garnett TO, Filippova M, Duerksen-hughes PJ. Accelerated degradation of FADD and procaspase 8 in cells expressing human papilloma virus 16 E6 impairs TRAIL-mediated apoptosis. Cell Death Differ, 2006, 13: 1915–1926.
Zhao LY, Colosimo AL, Liu Y, et al. Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcrip tion by Daxx. J Virol, 2003, 77: 11809–11821.
Song JJ, Lee YJ. Daxx deletion mutant (amino acids 501-625)-induced apoptosis occurs through the JNK/p38-Bax-dependent mitochondrial pathway. J Cell B iochem, 2004, 92: 1257–1270.
Yim EK, Lee KH, Bae J S, et al. Proteomic analysis of antiproliferative effects by treatment of 5-fluorouracil in cervical cancer cells. DNA Cell B iol, 2004, 23: 769–776.
Magal SS, Jackman A, Ish-Shalom S, et al. Downregulation of Bax mRNA expression and protein stability by the E6 protein of human papillomavirus 16. J Gen Virol, 2005, 86: 611–621.
Lee WT, Lee SH, Carriedo SG, et al. UV-vulnerability of human pap illoma virus type-16 E7-expressing astrocytes is associated with mitochondrial membrane depolarization and caspase-3 activation. Mol Cells, 2002, 14: 288–294.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supported by a grant from the education committee of Hunan province (No. 02C391).
Rights and permissions
About this article
Cite this article
He, A., Wang, X., Zhu, C. et al. Apoptosis of HeLa cells inhibited by human papillomavirus type 16 E6 protein interacting with hDaxx. Chin. -Ger. J. Clin. Oncol. 7, 728–731 (2008). https://doi.org/10.1007/s10330-008-0133-9
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10330-008-0133-9