Abstract
Squamous cell carcinoma (SCC) of the uterine cervix and oral cavity are most common cancers in India. Telomerase reverse transcriptase (TERT) overexpression is one of the hallmarks for cancer, and activation through promoter mutation C228T and C250T has been reported in variety of tumors and often shown to be associated with aggressive tumors. In the present study, we analyzed these two hot spot mutations in 181 primary tumors of the uterine cervix and oral cavity by direct DNA sequencing and correlated with patient’s clinicopathological characteristics. We found relatively high frequency of TERT hot spot mutations in both cervical [21.4 % (30/140)] and oral [31.7 % (13/41)] squamous cell carcinomas. In cervical cancer, TERT promoter mutations were more prevalent (25 %) in human papilloma virus (HPV)-negative cases compared to HPV-positive cases (20.6 %), and both TERT promoter mutation and HPV infection were more commonly observed in advanced stage tumors (77 %). Similarly, the poor and moderately differentiated tumors of the uterine cervix had both the TERT hot spot mutations and HPV (16 and 18) at higher frequency (95.7 %). Interestingly, we observed eight homozygous mutations (six 228TT and two 250TT) only in cervical tumors, and all of them were found to be positive for high-risk HPV. To the best of our knowledge, this is the first study from India reporting high prevalence of TERT promoter mutations in primary tumors of the uterine cervix and oral cavity. Our results suggest that TERT reactivation through promoter mutation either alone or in association with the HPV oncogenes (E6 and E7) could play an important role in the carcinogenesis of cervical and oral cancers.
Similar content being viewed by others
References
Ferlay JSI, Ervik M, Dikshit R, Eser S, Mathers C, Rebelo M, et al. GLOBOCAN 2012 v1.0, cancer incidence and mortality worldwide: IARC cancer base no. 11. Lyon: International Agency for Research on Cancer; 2012.
Ferlay J SI, Ervik M, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F. GLOBOCAN 2012 v1.0, cancer incidence and mortality worldwide: IARC cancer base no. 11. International Agency for Research on Cancer 1.0. 2013.
Bosch FX, de Sanjose S Chapter 1: human papillomavirus and cervical cancer—burden and assessment of causality. J Natl Cancer Inst Monogr. 2003;3–13.
Dikshit R, Gupta PC, Ramasundarahettige C, Gajalakshmi V, Aleksandrowicz L, et al. Cancer mortality in India: a nationally representative survey. Lancet. 2012;379:1807–16.
Byakodi R, Byakodi S, Hiremath S, Byakodi J, Adaki S, et al. Oral cancer in India: an epidemiologic and clinical review. J Community Health. 2012;37:316–9.
IARC Working Group on the Evaluation of Carcinogenic Risks to Humans. Betel-quid and areca-nut chewing and some areca-nut-derived nitrosamines. IARC monographs on the evaluation of carcinogenic risks to humans (2003). v. 85.
Gupta PC, Ray CS, Sinha DN, Singh PK. Smokeless tobacco: a major public health problem in the SEA region: a review. Indian J Public Health. 2011;55:199–209.
Wright WE, Piatyszek MA, Rainey WE, Byrd W, Shay JW. Telomerase activity in human germline and embryonic tissues and cells. Dev Genet. 1996;18:173–9.
Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, et al. Specific association of human telomerase activity with immortal cells and cancer. Science. 1994;266:2011–5.
Shay JW, Bacchetti S. A survey of telomerase activity in human cancer. Eur J Cancer. 1997;33:787–91.
Horn S, Figl A, Rachakonda PS, Fischer C, Sucker A, et al. TERT promoter mutations in familial and sporadic melanoma. Science. 2013;339:959–61.
Huang FW, Hodis E, Xu MJ, Kryukov GV, Chin L, et al. Highly recurrent TERT promoter mutations in human melanoma. Science. 2013;339:957–9.
Zhang Z, Yang X, Meng L, Liu F, Shen C, et al. Enhanced amplification of GC-rich DNA with two organic reagents. Biotechniques. 2009;47:775–9.
Vogelstein B, Kinzler KW. The path to cancer—three strikes and you’re out. N Engl J Med. 2015;373:1895–8.
Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144:646–74.
Cong YS, Wen J, Bacchetti S. The human telomerase catalytic subunit hTERT: organization of the gene and characterization of the promoter. Hum Mol Genet. 1999;8:137–42.
Liu X, Bishop J, Shan Y, Pai S, Liu D, et al. Highly prevalent TERT promoter mutations in aggressive thyroid cancers. Endocr Relat Cancer. 2013;20:603–10.
Rachakonda PS, Hosen I, de Verdier PJ, Fallah M, Heidenreich B, et al. TERT promoter mutations in bladder cancer affect patient survival and disease recurrence through modification by a common polymorphism. Proc Natl Acad Sci U S A. 2013;110:17426–31.
Killela PJ, Reitman ZJ, Jiao Y, Bettegowda C, Agrawal N, et al. TERT promoter mutations occur frequently in gliomas and a subset of tumors derived from cells with low rates of self-renewal. Proc Natl Acad Sci U S A. 2013;110:6021–6.
Griewank KG, Murali R, Schilling B, Schimming T, Moller I, et al. TERT promoter mutations are frequent in cutaneous basal cell carcinoma and squamous cell carcinoma. PLoS One. 2013;8:e80354.
Landa I, Ganly I, Chan TA, Mitsutake N, Matsuse M, et al. Frequent somatic TERT promoter mutations in thyroid cancer: higher prevalence in advanced forms of the disease. J Clin Endocrinol Metab. 2013;98:E1562–6.
Liu X, Wu G, Shan Y, Hartmann C, von Deimling A, et al. Highly prevalent TERT promoter mutations in bladder cancer and glioblastoma. Cell Cycle. 2013;12:1637–8.
Nault JC, Mallet M, Pilati C, Calderaro J, Bioulac-Sage P, et al. High frequency of telomerase reverse-transcriptase promoter somatic mutations in hepatocellular carcinoma and preneoplastic lesions. Nat Commun. 2013;4:2218.
Vinagre J, Almeida A, Populo H, Batista R, Lyra J, et al. Frequency of TERT promoter mutations in human cancers. Nat Commun. 2013;4:2185.
Cheng KA, Kurtis B, Babayeva S, Zhuge J, Tantchou I, et al. Heterogeneity of TERT promoter mutations status in squamous cell carcinomas of different anatomical sites. Ann Diagn Pathol. 2015;19:146–8.
Wu RC, Ayhan A, Maeda D, Kim KR, Clarke BA, et al. Frequent somatic mutations of the telomerase reverse transcriptase promoter in ovarian clear cell carcinoma but not in other major types of gynaecological malignancy. J Pathol. 2014;232:473–81.
Scott GA, Laughlin TS, Rothberg PG. Mutations of the TERT promoter are common in basal cell carcinoma and squamous cell carcinoma. Mod Pathol. 2014;27:516–23.
Mitra AB, Murty VV, Singh V, Li RG, Pratap M, et al. Genetic alterations at 5p15: a potential marker for progression of precancerous lesions of the uterine cervix. J Natl Cancer Inst. 1995;87:742–5.
Mitra AB, Murty VV, Li RG, Pratap M, et al. Allelotype analysis of cervical carcinoma. Cancer Res. 1994;54:4481–7.
Atkin NB. Significance of chromosome 5 and 17 changes in the development of carcinoma of the cervix uteri. Cytogenet Cell Genet. 2000;91:44–6.
Gewin L, Myers H, Kiyono T, Galloway DA. Identification of a novel telomerase repressor that interacts with the human papillomavirus type-16 E6/E6-AP complex. Genes Dev. 2004;18:2269–82.
Kiyono T, Foster SA, Koop JI, McDougall JK, Galloway DA, et al. Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cells. Nature. 1998;396:84–8.
Liu X, Dakic A, Chen R, Disbrow GL, Zhang Y, et al. Cell-restricted immortalization by human papillomavirus correlates with telomerase activation and engagement of the hTERT promoter by Myc. J Virol. 2008;82:11568–76.
Liu X, Disbrow GL, Yuan H, Tomaic V, Schlegel R. Myc and human papillomavirus type 16 E7 genes cooperate to immortalize human keratinocytes. J Virol. 2007;81:12689–95.
Liu X, Roberts J, Dakic A, Zhang Y, Schlegel R. HPV E7 contributes to the telomerase activity of immortalized and tumorigenic cells and augments E6-induced hTERT promoter function. Virology. 2008;375:611–23.
Liu X, Yuan H, Fu B, Disbrow GL, Apolinario T, et al. The E6AP ubiquitin ligase is required for transactivation of the hTERT promoter by the human papillomavirus E6 oncoprotein. J Biol Chem. 2005;280:10807–16.
Van Doorslaer K, Burk RD. Association between hTERT activation by HPV E6 proteins and oncogenic risk. Virology. 2012;433:216–9.
Oh ST, Kyo S, Laimins LA. Telomerase activation by human papillomavirus type 16 E6 protein: induction of human telomerase reverse transcriptase expression through Myc and GC-rich Sp1 binding sites. J Virol. 2001;75:5559–66.
Howie HL, Katzenellenbogen RA, Galloway DA. Papillomavirus E6 proteins. Virology. 2009;384:324–34.
Tungteakkhun SS, Duerksen-Hughes PJ. Cellular binding partners of the human papillomavirus E6 protein. Arch Virol. 2008;153:397–408.
Acknowledgments
We thank Dr. Sindhuja, Mrs. Arivazhagi (Arignar Anna Memorial Cancer Hospital & Research Institute, Kanchipuram), and Dr. V. Rajalakshmi (Institute of Obstetrics & Gynaecology and Government General Hospital for Women and Children, Chennai) for clinical assessment and sample collection. We, VV, KA, MM, AKD, and GA gratefully acknowledge the Government of India’s Council of Scientific and Industrial Research (CSIR) and University Grant Commission (UGC), respectively, for providing research fellowships. We also thank DST-FIST and UGC-SAP infrastructure facilities.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflicts of interest
None
Ethics approval
The Institutional Ethics Committee, Government Arignar Anna Memorial Cancer Hospital, Kancheepuram (No.101041/e1/2009-2), and the Madras Medical College, Chennai (No.04092010), approved the present study. Cervical and oral cancer samples were collected following the Institutional Ethical Committee (IEC) guidelines and informed consent was obtained from each patient, after explaining about the research study. For the illiterate patients, the study was verbally explained and consent was obtained with their thumb impression.
Additional information
Vilvanathan Vinothkumar and Ganesan Arunkumar contributed equally to this work.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Supplementary Table S1
Complete clinico-pathological profile of cervical cancer patients with HPV infection and TERT hot spot mutation status. (XLSX 15 kb)
Supplementary Table S2
Complete clinico-pathological profile of oral cancer patients with HPV infection and TERT hot spot mutation status. (XLSX 11 kb)
Rights and permissions
About this article
Cite this article
Vinothkumar, V., Arunkumar, G., Revathidevi, S. et al. TERT promoter hot spot mutations are frequent in Indian cervical and oral squamous cell carcinomas. Tumor Biol. 37, 7907–7913 (2016). https://doi.org/10.1007/s13277-015-4694-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13277-015-4694-2