Document Type : Original Research


1 Dept. of New Sciences and Technology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Dept. of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

3 Dept. of Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

4 Antimicrobial Resistance Research Center, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran


Background: Human papillomavirus (HPV) is responsible for the development of cervical neoplasia.  Infection with human papillomavirus type 16 (HPV-16) is a major risk factor for the development of cervical cancer. The virus encodes three oncoproteins (E5, E6 and E7), of which, the E7 oncoprotein is the major protein involved in cell immortalization and transformation of the infected cells. The aim of the current study was to develop Michigan Cancer Foundation 7 (MCF7) cells, which could stably express E7 protein of HPV type 16. Methods: E7 gene of HPV type 16 was introduced into MCF7 cells by Lipofectamine 2000 reagent and the transfected cells were treated with G418 antibiotic. After antibiotic selection of the transfected cells, stable expression of E7 gene of HPV16 was confirmed by reverse transcriptase polymerase chain reaction (RT-PCR). Results: Antibiotic selections of transfected cells were performed and transfected cells were alive in cytotoxic concentration of the antibiotic. RNA was extracted from transfected cells and E7 gene of HPV16 was amplified by RT-PCR method and a 350-bp band corresponds to E7 was observed. Conclusion: Results confirmed the stable transfection of cells. The stably transfected cells can be used as a useful tool in future studies on HPV16 and cancers caused by this virus.


How to cite this article:
Ghasemi F, Rostami S, Sadat Nabavinia M, Meshkat Z. Developing Michigan Cancer Foundation 7 Cells with Stable Expression of E7 Gene of Human Papillomavirus Type 16. Iran J Pathol. 2016; 11(1):41-46.


Main Subjects

  1. Walboomers JM, Jacobs MV, Manos MM, Bosch FX, Kummer JA, Shah KV, et al. Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 1999;189(1):12-9.
  2. Bosch FX, Manos MM, Muñoz N, Sherman M, Jansen AM, Peto J, et al. Prevalence of human papillomavirus in cervical cancer: a worldwide perspective. J Natl Cancer Inst 1995;87(11):796-802.
  3. Fernandes J, Araújo J, Fernandes T. Biology and natural history of human papillomavirus infection. Open J Clin Trials 2013;5:1-12.
  4. Koutsky P, Laura. Epidemiology of genital human papillomavirus infection. Am J Med 1997;102(5S1):3-8.
  5. Winer RL, Lee S-K, Hughes JP, Adam DE, Kiviat NB, Koutsky LA. Genital human papillomavirus infection: incidence and risk factors in a cohort of female university students. Am J Epidemiol 2003;157(3):218-26.
  6. Khan MJ, Castle PE, Lorincz AT, Wacholder S, Sherman M, Scott DR, et al. The elevated 10-year risk of cervical precancer and cancer in women with human papillomavirus (HPV) type 16 or 18 and the possible utility of type-specific HPV testing in clinical practice. J Natl Cancer Inst 2005;97(14):1072-9.
  7. Liaw K-L, Hildesheim A, Burk RD, Gravitt P, Wacholder S, Manos MM, et al. A prospective study of human papillomavirus (HPV) type 16 DNA detection by polymerase chain reaction and its association with acquisition and persistence of other HPV types. J Infect Dis 2001;183(1):8-15.
  8. Ghittoni R, Accardi R, Hasan U, Gheit T, Sylla B, Tommasino M. The biological properties of E6 and E7 oncoproteins from human papillomaviruses. Virus Genes 2010;40(1):1-13.
  9. Marur S, D'Souza G, Westra WH, Forastiere AA. HPV-associated head and neck cancer: a virus-related cancer epidemic. Lancet Oncol 2010;11(8):781-9.
  10. Nichols AC, Dhaliwal SS, Palma DA, Basmaji J, Chapeskie C, Dowthwaite S, et al. Does HPV type affect outcome in oropharyngeal cancer? J Otolaryngol Head Neck Surg 2013;42(1):1-9.
  11. Hanahan D, Weinberg R. Hallmarks of cancer: the next generation. Cell 2011;144(5):646-74.
  12. Meshkat Z, Soleimanjahi H, Mahmoudi M, Hassan ZM, Mirshahabi H, Meshkat M, et al. CTL Responses to a DNA Vaccine Encod-ing E7 Gene of Human Papillomavirus Type 16 from an Iranian Isolate. Iran J Immunol 2008;5(2):82-91.
  13. Villa LL, Costa RL, Petta CA, Andrade RP, Ault KA, Giuliano AR, et al. Prophylactic quadrivalent human papillomavirus (types 6, 11, 16, and 18) L1 virus-like particle vaccine in young women: a randomised double-blind placebo-controlled multicentre phase II efficacy trial. Lancet Oncol 2005;6(5):271-8.
  14. Clifford G, Franceschi S, Diaz M, Munoz N, Villa LL. HPV type-distribution in women with and without cervical neoplastic diseases. Vaccine 2006;24:S26-S34.
  15. Bresse X, Adam M, Largeron N, Roze S, Marty R. A comparative analysis of the epidemiological impact and disease cost-savings of HPV vaccines in France. Hum Vaccin Immunother 2013;9(4).
  16. Petry KU, Luyten A, Justus A, Iftner A, Strehlke S, Reinecke-Lüthge A, et al. Prevalence of high-risk HPV types and associated genital diseases in women born in 1988/89 or 1983/84--results of WOLVES, a population-based epidemiological study in Wolfsburg, Germany. BMC Infect Dis 2013;13(1):135.
  17. Pontén J, Adami H, Friberg L, Gustafsson L, Miller A, Parkin M, et al. HPV and cervical cancer. Int J Cancer 1995;63(2):317-20.
  18. Edwards TG, Helmus MJ, Koeller K, Bashkin JK, Fisher C. Human Papillomavirus Episome Stability Is Reduced by Aphidicolin and Controlled by DNA Damage Response Pathways. J Virol 2013;87(7):3979-89.
  19. DeFilippis RA, Goodwin EC, Wu L, DiMaio D. Endogenous human papillomavirus E6 and E7 proteins differentially regulate proliferation, senescence, and apoptosis in HeLa cervical carcinoma cells. J Virol 2003;77(2):1551-63.
  20. Teissier S, Pang C, Thierry F. The E2F5 repressor is an activator of E6/E7 transcription and of the S-phase entry in HPV18-associated cells. Oncogene 2010;29(36):5061-70.
  21. Morandell D, Kaiser A, Herold S, Rostek U, Lechner S, Mitterberger MC, et al. The human papillomavirus type 16 E7 oncoprotein targets Myc-interacting zinc-finger protein-1. Virology 2012;422(2):242-53.
  22. Magaldi TG, Almstead LL, Bellone S, Prevatt EG, Santin AD, DiMaio D. Primary human cervical carcinoma cells require human papillomavirus E6 and E7 expression for ongoing proliferation. Virology 2012;422(1):114-24.
  23. Ling M, Wang X, Tsao S, Wong Y. Down-regulation of Id-1 expression is associated with TGFβ1-induced growth arrest in prostate epithelial cells. Biochim Biophys Acta 2002;1570(3):145-52.
  24. Akil N, Yasmeen A, Kassab A, Ghabreau L, Darnel A, Al Moustafa A. High-risk human papillomavirus infections in breast cancer in Syrian women and their association with Id-1 expression: a tissue microarray study. Br J Cancer 2008;99(3):404-7.
  25. Arroyo M, Bagchi S, Raychaudhuri P. Association of the human papillomavirus type 16 E7 protein with the S-phase-specific E2F-cyclin A complex. Mol cell Biol 1993;13(10):6537-46.
  26. Li F. Role of survivin and its splice variants in tumorigenesis. Br J Cancer 2004;92(2):212-6.
  27. Borbély ÁA, Murvai M, Kónya J, Beck Z, Gergely L, Li F, et al. Effects of human papillomavirus type 16 oncoproteins on survivin gene expression. J Gen Virol 2006;87(2):287-94.
  28. Yan J, Yong W, Lei XIONG YY, Jia Rui W. Differential regulation of survivin by p53 contributes to cell cycle dependent apoptosis. Cell Res 2005;15(5):361-70.