Abstract
Human papillomavirus (HPV) infection has been identified as an etiologic factor of head and neck cancers (HNCs). We explored the potential use of antisense HPV RNA transcripts for gene therapy and its effect in combination with cisplatin (CDDP) for HPV-positive HNCs. We introduced the antisense RNA transcripts of the E6 and E7 genes of HPV type 16 into UM-SCC-47 cells harboring HPV 16 and YCU-T892 cells that were HPV-negative using a recombinant adenoviral vector, Ad-E6/E7-AS. We then analyzed the effects of the introduction of Ad-E7-AS on cell and tumor growth and the synergistic effect with CDDP in vitro and in vivo. After infection of Ad-E6/E7-AS, the cellular growth of UM-SCC-47 cells were suppressed, but not that of YCU-T892 cells. E7 protein expression was suppressed, and p53 and pRb protein expression increased after infection of Ad-E7-AS. Cell growth and tumorigenicity were greatly suppressed in combination with CDDP compared with Ad-E7-AS or CDDP treatment alone in vitro. Ad-E7-AS combined with CDDP treatment significantly reduced the volumes of established subcutaneous tumors. Transfection with HPV 16 E7 antisense RNA combined with CDDP treatment might be a potentially useful approach to the therapy of HPV 16-positive HNC.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Global Burden of Disease Cancer C, Fitzmaurice C, Allen C, Barber RM, Barregard L, Bhutta ZA, et al. Global, regional, and national cancer incidence, mortality, years of life lost, years lived with disability, and disability-adjusted life-years for 32 cancer groups, 1990 to 2015: a systematic analysis for the global burden of disease study. JAMA Oncol. 2017;3:524e48.
UICC International Union Against Cancer. HPV mediated oropharyngeal cancer. TNM classification of malignant tumours. In: Brierley JD, Gospodarowicz MK, Wittekind,Ch.10. Wiley-Blackwell; 2016. p. 113–122.
Fakhry C, Westra WH, Li S, Cmelak A, Ridge JA, Pinto H, et al. Improved survival of patients with human papillomavirus-positive head and neck squamous cell carcinoma in a prospective clinical trial. J Natl Cancer Inst. 2008;100:261e9.
Chen AM, Felix C, Wang PC, Hsu S, Basehart V, Garst J, et al. Reduced-dose radiotherapy for human papillomavirus-associated squamous-cell carcinoma of the oropharynx: a single-arm, phase 2 study. Lancet Oncol. 2017;18:803–11.
Masterson L, Moualed D, Liu ZW, Howard JE, Dwivedi RC, Tysome JR, et al. De-escalation treatment protocols for human papillomavirus-associated oropharyngeal squamous cell carcinoma: a systematic review and meta-analysis of current clinical trials. Eur J Cancer. 2014;50:2636–48.
Trotti A. Toxicity in head and neck cancer: a review of trends and issues. Int J Radiat Oncol. 2015;47:1–12.
Machtay M, Moughan J, Trotti A, Garden AS, Weber RS, Cooper JS, et al. Factors associated with severe late toxicity after concurrent chemoradiation for locally advanced head and neck cancer: an RTOG analysis. J Clin Oncol. 2008;26:3582–9.
Ndiaye C, Mena M, Alemany L, Arbyn M, Castellsague X, Laporte L, et al. HPV DNA, E6/E7 mRNA, and p16INK4a detection in head and neck cancers: a systematic review and meta-analysis. Lancet Oncol. 2014;15:1319e31.
Bradford CR, Zhu S, Ogawa H, Ogawa T, Ubell M, Narayan A, et al. P53 mutation correlates with cisplatin sensitivity in head and neck squamous cell carcinoma lines. Head Neck. 2003;25:654–61.
Adhim Z, Lin X, Huang W, Morishita N, Kawabata M, Nakamura T, Yasui H, et al. E10A, an adenovirus-carrying endostatin gene, dramatically increased the tumor drug concentration of metronomic chemotherapy with low-dose cisplatin in a xenograft mouse model for head and neck squamous-cell carcinoma. Cancer Gene Ther. 2012;19:144–52.
Tan TM, Ting RC. In vitro and in vivo inhibition of human papillomavirus type 16 E6 and E7 genes. Cancer Res. 1995;55:4599–605.
Chen Z, Kamath P, Zhang S, St John L, Adler-Storthz K, Shillitoe EJ. Effects on tumor cells of ribozymes that cleave the RNA transcripts of human papillomavirus type 18. Cancer Gene Ther. 1996;3:18–23.
Sima N, Wang S, Wang W, Kong D, Xu Q, Tian X, et al. Antisense targeting human papillomavirus type 16 E6 and E7 genes contributes to apoptosis and senescence in SiHa cervical carcinoma cells. Gynecol Oncol. 2007;106:299–304.
Jonson AL, Rogers LM, Ramakrishnan S, Downs LS Jr.. Gene silencing with siRNA targeting E6/E7 as a therapeutic intervention in a mouse model of cervical cancer. Gynecol Oncol. 2008;111:356–64.
Adhim Z, Otsuki N, Kitamoto J, Morishita N, Kawabata M, Shirakawa T, et al. Gene silencing with siRNA targeting E6/E7 as a therapeutic intervention against head and neck cancer-containing HPV16 cell lines. Acta Otolaryngol. 2013;133:761–71.
Smotkin D, Prokoph H, Wettstein FO. Oncogenic and nonooncigenic human genital papillomaviruses generate theE7 mRNA by different mechanisms. J Virol. 1989;63:1441–7.
Butz K, Ristriani T, Hengstermann A, Denk C, Scheffner M, Hoppe-Seyler F. siRNA targeting of the viral E6 oncogene efficiently kills human papillomavirus-positive cancer cells. Oncogene. 2003;22:5938–45.
Ziemann F, Arenz A, Preising S, Wittekindt C, Klussmann JP, Engenhart-Cabillic R, et al. Incresed sensitivity of HPV-positive head and neck cancer cell lines to x-irradiation + Cisplatin due to decreased expression of E6 and E7 oncoproteins and enhanced apoptosis. Am J Cancer Res. 2015;5:1017–31.
Li L, Cui Xu, Jia Long, Shen D, Zhou W, Zhou Q, et al. E7 and E7 gene silencing results in decreased methylation of tumor suppressor genes and induces phenotype transformation of human cervical carcinoma cell lines. Oncotarget. 2015;6:23930–43.
Putral LN, Bywater MJ, Sauders NA, Gabrielli BG, Leggatt GR, Mcmillan NAJ. RNA interference against himan papillomavirus oncogenes in cervical cancer cells results in increased sensitivity to cisplatin. Mol Pharmacol. 2005;68:1311–19.
Jung HS, Erkin AC, Kwon MJ, Kim AH, Jung JI, Oh YK, et al. The synergic therapeutic effect of cisplatin with human papillomavirus E6/E7 short interfering RNA on cervical cancer cell lines in vitro and in vivo. Int J Cancer. 2012;130:1925–36.
Koivusalo R, Krausz E, Helenius H, Hietanen S. Chemotherapy compounds in cervical cancer cells primed by reconstitution of p53 function after short interfering RNA-mediated degradation of human papillomavirus 18 E6 mRNA: opposite effect of siRNA in combination with different drugs. Mol Pharmacol. 2005;68:372–82.
Acknowledgements
We express our gratitude to Dr. Thomas E. Carey and Dr. Carol R. Bradford, University of Michigan, for kindly providing the UM-SCC-47 cell lines, and M. Kurihara for assisting immunohistochemical analyses. This research was supported by Grants-in-aid for Scientific Research (C) from Japan Society for the Promotion of Science: project number 25462683 (Otsuki).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Kojima, Y., Otsuki, N., Kubo, M. et al. Adenovirus-mediated transfer of HPV 16 E6/E7 antisense RNA combined with cisplatin inhibits cellular growth and induces apoptosis in HPV-positive head and neck cancer cells. Cancer Gene Ther 25, 274–283 (2018). https://doi.org/10.1038/s41417-018-0024-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41417-018-0024-3
This article is cited by
-
Evaluation of HPV16 E7 expression in head and neck carcinoma cell lines and clinical specimens
Scientific Reports (2020)