DOI QR코드

DOI QR Code

Promoter Methylation and Relative mRNA Expression of the p16 Gene in Cervical Cancer in North Indians

  • Gupta, Amita (Department of Obstetrics and Gynaecology, King George's Medical University) ;
  • Ahmad, Mohammad Kaleem (Department of Biochemistry, King George's Medical University) ;
  • Mahndi, Abbas Ali (Department of Biochemistry, King George's Medical University) ;
  • Singh, Renu (Department of Obstetrics and Gynaecology, King George's Medical University) ;
  • Pradeep, Yashodhara (Department of Obstetrics and Gynaecology, King George's Medical University)
  • 발행 : 2016.08.01

초록

Background: Cervical carcinoma is one of the main causes of mortality in women worldwide as well as in India. It occurs as a result of various molecular events that develop from the combined influences of an individual's genetic predisposition and external agents such as smoking and menstrual hygiene, for example. However, infection with human papillomavirus (HPV) is the established major risk factor. The aim of the current study was to investigate p16 CpG island methylation and establish any correlation with mRNA expression in north Indian population. Materials and Methods: We analyzed 196 woman volunteer out of which 98 were cases and 98 healthy controls. For the analysis of methylation pattern, DNA extracted from blood samples was modified with a bisulfate kit and used as template for methylation specific PCR (MSP). Quantitative real-time PCR (QRT-PCR) was performed to check mRNA expression. Results: Correlation between methylation status of p16 gene and poor menstrual hygiene was significant (p=0.006), high parity cases showed methylation of p16 gene (p=0.031) with increased risk up to 1.86 times for cervical cancer and smoking was a strong risk factor associated with cervical cancer. We analyzed methylation pattern and found 60.3% methylation in cases with low mRNA expression level (0.014) as compare to controls (1.24). It was also observed that promoter methylation of p16 gene was significantly greater in FIGO stage III. Conclusions: We conclude that p16 methylation plays an important role in cervical cancer in the north Indian population and its methylation decreases mRNA expression. It can be used as an important and consistent blood biomarker in cervical cancer patients.

키워드

참고문헌

  1. Akintola AD, Crislip ZL, Catania JM, et al (2008). Promoter methylation is associated with the age-dependent loss of N-cadherin in the rat kidney. Am J Renal Physiol, 294, 170-6. https://doi.org/10.1152/ajprenal.00285.2007
  2. Appleby P, Beral V, Berrington de Gonzalez A, et al (2007). Cervical Cancer and hormonal contraceptives: collaborative reanalysis of individual data for 16,573 women with cervical cancer and 35,509 women without cervical cancer from 24 epidemiological studies. Lancet, 370, 1609-21. https://doi.org/10.1016/S0140-6736(07)61684-5
  3. Au WW, Abdou-Salama S, Sierra-Torres CH, et al (2007). Environmental risk factors for prevention and molecular intervention of cervical cancer. Int J Hyg Environ Health, 210, 671-8. https://doi.org/10.1016/j.ijheh.2006.10.003
  4. Bayo S, Bosch FX, de Sanjose S, et al (2002). Risk factors of invasive cervical cancer in Mali. Int J Epidemiol, 31, 202-9. https://doi.org/10.1093/ije/31.1.202
  5. Bosch FX, Manos MM, Muñoz N, et al (1995). Prevalence of Human Papillomavirus in Cervical Cancer: a worldwide perspective. International biological study on cervical cancer (IBSCC) Study Group. J Natl Cancer Inst, 87, 796-02. https://doi.org/10.1093/jnci/87.11.796
  6. Brinton LA, Schairer C, Haenszel W, et al (1986). Cigarette smoking and invasive cervical cancer. JAMA, 255, 3265-9. https://doi.org/10.1001/jama.1986.03370230071033
  7. Cao J, Zhou J, Gao Y, et al (2009). Methylation of p16 CpG islands associated with malignant progression of oral epithelial dysplasia: a prospective cohort study. Clin Cancer Res, 15, 5178-83. https://doi.org/10.1158/1078-0432.CCR-09-0580
  8. Chomczynski P, Sacchi N (1987). Single-step method of RNA isolation by acid guanidiniumthiocyanate-phenolchloroform extraction. Anal Biochem, 162, 156-9.
  9. Dillner J, Lehtinen M, Bjorge T, et al (1997). Prospective seroepidemiologic study of human papillomavirus infection as a risk factor for invasive cervical cancer. J Natl Cancer Inst, 89, 1293-99. https://doi.org/10.1093/jnci/89.17.1293
  10. Esteller M (2002). CpG island hypermethylation and tumor suppressor genes: a booming present, a brighter future. Oncogene, 21, 5427-40. https://doi.org/10.1038/sj.onc.1205600
  11. Franceschi S, Rajkumar T, Vaccarella S, et al (2003). Human papillomavirus and risk factors for cervical cancer in Chennai, India: a case-control study. Int J Cancer, 107, 127-33. https://doi.org/10.1002/ijc.11350
  12. GLOBOCAN (2013). World Health Organization.
  13. Gupta MK, Singh R, Banerjee M (2016). Cytokine gene polymorphisms and their association with cervical cancer: A North Indian study. Egypt J Med Hum Genet, 17, 155-63. https://doi.org/10.1016/j.ejmhg.2015.10.005
  14. Hanahan D, Weinberg RA (2000). The hallmarks of cancer. Cell, 100, 57-70. https://doi.org/10.1016/S0092-8674(00)81683-9
  15. Herman JG, Graff JR, Myohanen S, et al (1996). Methylation specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci, 93, 9821-6. https://doi.org/10.1073/pnas.93.18.9821
  16. Hibi K, Taguchi M, Nakayama H, et al (2001). Molecular detection of p16 promoter methylation in the serum of patients with esophageal squamous cell carcinoma. Clin Cancer Res, 7, 3135-8.
  17. Hildesheim A, Herrero R, Castle PE, et al (2001). HPV co-factors related to the development of cervical cancer: results from a population-based study in Costa Rica. Br J Cancer, 84, 1219-26. https://doi.org/10.1054/bjoc.2001.1779
  18. Kjellberg L, Hallmans G, Ahren AM, et al (2000). Smoking, diet, pregnancy and oral contraceptive use as risk factors for cervical intra-epithelial neoplasia in relation to human papillomavirus infection. Br J Cancer, 82, 1332-8. https://doi.org/10.1054/bjoc.1999.1100
  19. Kruger-Kjaer S, van den Brule AJ, Svare EI, et al (1998). Different risk factor patterns for high-grade and low-grade intraepithelial lesions on the cervix among HPV-positive and HPV-negative young women. Int J Cancer, 76, 613-9. https://doi.org/10.1002/(SICI)1097-0215(19980529)76:5<613::AID-IJC1>3.0.CO;2-T
  20. Liu H, Liu XW, Dong G, et al (2015). P16 methylation as an early predictor for cancer development from oral epithelial dysplasia: a double-blind multicentre prospective study. E Bio Medicine, 2, 432-7.
  21. Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative pcr and the 2− ${\delta}{\delta}$ct method. Methods, 25, 402-8. https://doi.org/10.1006/meth.2001.1262
  22. Louie KS, Castellsague X, de Sanjose S, et al (2011). Smoking and passive smoking in cervical cancer risk: pooled analysis of couples from the IARC multicentric case-control studies. Cancer Epidemiol Biomarkers Prev, 20, 1379-90. https://doi.org/10.1158/1055-9965.EPI-11-0284
  23. Luo D, Zhang B, Lv L, et al (2006). Methylation of CpG islands of p16 associated with progression of primary gastric carcinomas. Lab Invest, 86, 591-8. https://doi.org/10.1038/labinvest.3700415
  24. Marjani HA, Biramijamal F, Rakhshani N, et al (2010). Investigation of NQO1 genetic polymorphism, NQO1 gene expression and PAH-DNA adducts in ESCC.A case-control study from Iran. Genet Mol Res, 9, 239-49. https://doi.org/10.4238/vol9-1gmr693
  25. Miller SA, Dykes DD, Polesky HF (1988). A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res, 16, 1215. https://doi.org/10.1093/nar/16.3.1215
  26. Mitra S (2009). Study of the risk factors for cancer cervix in a speciality hospital in Kolkata. J Com Med, 5, 1-5.
  27. National Comprehensive Cancer Network (NCCN)-Clinical Practice Guidelines in Oncology (2013). Cervical Cancer, 3.
  28. Nuovo GJ, Plaia TW, Belinsky SA, et al (1999). In situ detection of the hypermethylation-induced inactivation of the p16 gene as an early event in oncogenesis. Proc Natl Acad Sci, 96, 12754-9. https://doi.org/10.1073/pnas.96.22.12754
  29. Parkin DM, Bray F (2006). The burden of HPV- related cancers. Vaccine, 24, 11-25.
  30. Pecorelli S, Zigliani L, Odicino F (2009). Revised FIGO staging for carcinoma of the vulva, cervix, and endometrium. Int J Gynaecol Obstet, 105, 103-4. https://doi.org/10.1016/j.ijgo.2009.02.012
  31. Raychaudhuri S, Mandal S (2012). Socio-demographic and behavioural risk factors for cervical cancer and knowledge, attitude and practice in rural and urban areas of North Bengal, India. Asian Pac J Cancer Prev, 4, 1093-6.
  32. Sharma S, Kelly TK, Jones PA (2010). Epigenetics in cancer. Carcinogenesis, 31, 27-36. https://doi.org/10.1093/carcin/bgp220
  33. Shim YH, Park HJ, Choi MS, et al (2003). Hypermethylation of the p16 gene and lack of p16 expression in hepatoblastoma. Mod Pathol, 16, 430-6. https://doi.org/10.1097/01.MP.0000066799.99032.A7
  34. Vaccarella S, Herrero R, Dai M, et al (2006). Reproductive factors, oral contraceptive use, and human papillomavirus infection: pooled analysis of the IARC HPV prevalence surveys. Cancer Epidemiol Biomarkers Prev, 15, 2148-53. https://doi.org/10.1158/1055-9965.EPI-06-0556
  35. Virmani AK, Muller C, Rathi A, et al (2001). Aberrant methylation during cervical carcinogenesis. Clin Cancer Res, 7, 584-9.
  36. Weber M, Hellmann I, Stadler MB, et al (2007). Distribution, silencing potential and evolutionary impact of promoter DNA methylation in the human genome. Nat Genet, 39, 457-66. https://doi.org/10.1038/ng1990
  37. Weinberg RA (1995). The retinoblastoma protein and cell cycle control. Cell, 81, 323-30. https://doi.org/10.1016/0092-8674(95)90385-2