Association of Peripheral Blood RASSF1A and CDKN2A Methylation Status with Smoking Behaviour in Nasopharyngeal Carcinoma

Erika Diana Risanti, Aditya Kurniawan, Laila Wahyuningsih, Ery Kus Dwianingsih, Hanggoro Tri Rinonce, Jajah Fachiroh

Abstract


BACKGROUND: Hypermethylation of RASSF1A and CDKN2A is one of epigenetic factor underlies nasopharyngeal carcinoma (NPC) development. Smoking behavior as an NPC’s risk factor causes aberrant DNA methylation. RASSF1A and CDKN2A promoter hypermethylation from peripheral blood cells correlates with smoking behavior. The use of body fluids including peripheral blood as a specimen for DNA methylation analyzes are widely developed, as less invasive method compared to the use of tissue biopsy. This study aims to observe the association between RASSF1A and CDKN2A methylation in peripheral blood and smoking behavior
among NPC patients.

METHODS: Newly diagnosed NPC subjects were recruited from ear-nose-throat (ENT) outpatient clinic of Dr. Sardjito Hospital, Yogyakarta. DNA from buffycoat of 19 smokers and 20 non-smokers NPC’s patients were isolated. Bisulphite modification was applied to 500 ng of the isolated DNA. The methylation status was detected by MSP (methylation-specific polymerase chain reaction (PCR)). The association between smoking status and promoter hypermethylation was analysis using Chi-Square test.

RESULTS: MSP analysis of RASSF1A showed that 68.42% smoker and 75% non-smoker NPC’s patients were methylated. MSP analysis of CDKN2A showed that 21.05% smoker and 25% non-smoker NPC’s patients were methylated. There was no association between smoking behavior with RASSF1A and CDKN2A methylation (p>0.05).

CONCLUSION: Statistical analysis showed that smoking behavior is not associated with methylation of RASSF1A and CDKN2A among NPC’s patients.

KEYWORDS: DNA methylation, CDKN2A, RASSF1A, Nasopharyngeal carcinoma, Smoking


Full Text:

PDF

References


Tsao SW, Yip YL, Tsang CM, Pang PS, Lau VMY, Zhang G, et al. Etiological factors of nasopharyngeal carcinoma. Oral Oncol. 2014; 50: 330-8, CrossRef.

Adham M, Middeldorp JM, Kurniawan AN, Muhtadi AI, Roezin A, Hermani B, et al. Nasopharyngeal carcinoma in Indonesia: epidemiology, incidence, signs, and symptoms at presentation. Chin J Cancer. 2013; 31: 185-96, CrossRef.

Thompson LDR. Update on Nasopharyngeal Carcinoma. Head Neck Pathol. 2007; 1: 81-6, CrossRef.

Chou J, Ling YC, Kim J, You L, Xu Z, He B, et al. Nasopharyngeal carcinoma review of molecular mechanisms of tumorigenesis. Head Neck Cancer. 2008; 30: 946-63, CrossRef.

Lee KWK, Pausova Z. Cigarette smoking and DNA methylation. Front Genet. 2013; 4: 132, CrossRef.

Yokoi K, Yamashita K, Watanabe M. Analysis of DNA methylation status in bodily fluids for early detection of cancer. Int J Mol Sci. 2017; 18: E735, CrossRef.

Dai W, Zheng H, Cheung AKL, Lung ML. Genetic and epigenetic landscape of nasopharyngeal carcinoma. Chinese Clin Oncol. 2016; 5: 16, CrossRef.

Hutajulu SH, Indrasari SR, Indrawati LPL, Harijadi A, Duin S, Haryana SM, et al. Epigenetic markers for early detection of nasopharyngeal carcinoma in a high risk population. Mol Cancer. 2011; 10: 48, CrossRef.

Tian F, Yip SP, Kwong DLW, Lin Z, Yang Z, Wu VWC. Promoter hypermethylation of tumor suppressor genes in serum as potential biomarker for the diagnosis of nasopharyngeal carcinoma. Cancer Epidemiol. 2013; 37: 708-13, CrossRef.

Challouf S, Ziadi S, Zaghdoudi R, Ksiaa F, Ben Gacem R, Trimeche M. Patterns of aberrant DNA hypermethylation in nasopharyngeal carcinoma in Tunisian patients. Clin Chim Acta. 2012; 413: 795-802, CrossRef.

Huang T, Chen X, Hong Q, Deng Z, Ma H, Xin Y, et al. Meta-analyses of gene methylation and smoking behavior in non-small cell lung cancer patients. Sci Rep. 2015; 5: 8897, CrossRef.

Woodson K, Mason J, Choi S, Woodson K, Mason J, Choi S, et al. Hypomethylation of p53 in peripheral blood DNA is associated with the development of lung cancer hypomethylation of p53 in peripheral blood DNA is associated with the development of lung cancer. 2001; 10: 69-74, PMID.

Pedersen KS, Bamlet WR, Oberg AL, de Andrade M, Matsumoto ME, Tang H, et al. Leukocyte DNA methylation signature differentiates pancreatic cancer patients from healthy controls. PLoS One. 2011; 6: 1-9, CrossRef.

Teschendorff AE, Menon U, Gentry-Maharaj A, Ramus SJ, Gayther SA, Apostolidou S, et al. An epigenetic signature in peripheral blood predicts active ovarian cancer. PLoS One. 2009; 4: e8274, CrossRef.

Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci. 1996; 93: 9821-6, CrossRef.

Lo K, Kwong J, Hui AB, Chan SY, To K, Chan AS, et al. High frequency of promoter hypermethylation of RASSF1A in nasopharyngeal carcinoma. Cancer Res. 2001; 61: 3877-81, PMID.

Li L, Choi JY, Lee KM, Sung H, Park SK, Oze I, et al. DNA methylation in peripheral blood: a potential biomarker for cancer molecular epidemiology. J Epidemiol. 2012; 22: 384-94, CrossRef.

Walton E, Hass J, Liu J, Roffman JL, Bernardoni F, Roessner V, et al. Correspondence of DNA methylation between blood and brain tissue and its application to schizophrenia research. Schizophr Bull. 2016; 42: 406-14, CrossRef.

Li X, Wang Y, Zhang Z, Yao X, Ge J, Zhao Y. Correlation of MLH1 and MGMT methylation levels between peripheral blood leukocytes and colorectal tissue DNA samples in colorectal cancer patients. Oncol Lett. 2013; 6: 1370-6, CrossRef.

Deep JS, Sidhu S, Chandel A, Thapliyal S, Garg C. Aberrant methylation in promoters of GSTP1, p16, p14,and RASSF1A genes in smokers of North India. ISRN Pulmonol. 2012; 2012: 247631, CrossRef.




DOI: https://doi.org/10.18585/inabj.v10i2.381

Indexed by:

                 

                

                

  

 

The Prodia Education and Research Institute