Abstract
An optimized recombinant HPV16 E6E7 fusion gene (HPV16 ofE6E7) was constructed according to codon usage for mammalian cell expression, and a mutant of HPV16 ofE6E7 fusion gene (HPV16 omfE6E7) was generated by site-directed mutagenesis at L57G, C113R for the E6 protein and C24G, E26G for the E7 protein for HPV16 ofE6E7 [patent pending (CN 101100672)]. The HPV16 omfE6E7 gene constructed in this work not only lost the transformation capability to NIH 3T3 cells and tumorigenicity in SCID mice, but also maintained very good stability and antigenicity. These results suggests that the HPV16 omfE6E7 gene should undergo further study for application as a safe antigen-specific therapeutic vaccine for HPV16-associated tumors.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bafna S, Singh A P, Moniaux N, et al. 2008. MUC4, a multifunctional transmembrane glycoprotein, induces oncogenic transformation of NIH3T3 mouse fibroblast cells. Cancer Res, 68(22): 9231–9238.
Bequet-Romero M, Lopez-Ocejo O. 2000. Angiogenesis modulators expression in culture cell lines positives for HPV-16 oncoproteins. Biochem Biophys Res Commun, 277(1):55–61.
Edmonds C, Vousden K H. 1989. A point mutational analysis of human papillomavirus type 16 E7 protein. J Virol, 63(6): 2650–2656.
Hung C F, Monie A, Weng W H, et al. 2010. DNA vaccines for cervical cancer. Am J Transl Res, 2(1): 75–87.
Lopez-Ocejo O, Viloria-Petit A, Bequet-Romero M, et al. 2000. Oncogenes and tumor angiogenesis: the HPV-16 E6 oncoprotein activates the vascular endothelial growth factor (VEGF) gene promoter in a p53 independent manner. Oncogene, 19: 4611–4620.
Nadagawa S, Watanabe S, Yoshikawa H, et al. 1995. Mutational Analysis of Human Papillomavirus Type 16 E6 Protein: Transforming Function for Human Cells and Degradation of p53 in vitro. Virology, 212(2): 535–542.
Ohlschläger P, Quetting M, Alvarez G, et al. 2009. Enhancement of immunogenicity of a therapeutic cervical cancer DNA-based vaccine by co-application of sequenceoptimized genetic adjuvants. Int J Cancer, 125(1): 189–198.
Toussaint-Smith E, Donner D B, Roman A. 2004. Expression of human papillomavirus type 16 E6 and E7 oncoproteins in primary foreskin keratinocytes is sufficient to alter the expression of angiogenic factors. Oncogene, 23:2988–2995.
Walboomers J M, Jacobs M V, Manos M M, et al. 1999. Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol, 189: 12–19.
Watanabe S, Kanda T, Sato H, et al. 1990. Mutational analysis of human papillomavirus type 16 E7 functions. J Virol, 64(1): 207–214.
Yan J, Reichenbach D K, Corbitt N, et al. 2009. Induction of antitumor immunity in vivo following delivery of a novel HPV-16 DNA vaccine encoding an E6/E7 fusion antigen. Vaccine, 27(3): 431–440.
Zhou X S, Qian X L, Zhao Q Z, et al. 2004. Efficient expression of modified human papillomavirus 16 E6/E7 fusion protein and the antitumor efficacy in a mouse model. Biol Pharm Bull. 27(3): 303–307.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Foundation items: National 863 Program (2007AA 021107); Beijing Municipal Commission of Education Research Projects (KM201010005008); Doctoral Startup Foundation of Beijing University of Technology.
Rights and permissions
About this article
Cite this article
Xie, Q., Zhou, Zx., Li, Zl. et al. Transforming activity of a novel mutant of HPV16 E6E7 fusion gene. Virol. Sin. 26, 206–213 (2011). https://doi.org/10.1007/s12250-011-3178-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12250-011-3178-9