Promoter hypermethylation plays an important role in the inactivation of tumor suppressor/metabolic genes during tumorigenesis. The screening of high-risk population (smokers) for hypermethylation pattern in tumor suppressor/metabolic genes can be a good noninvasive biomarker tool, which should be included in prognosis so that therapeutic measures can be initiated at an early stage. The purpose of this study was to determine the prevalence of aberrant promoter methylation of GSTP1, p16, p14, and RASSF1A genes in smokers and nonsmokers of North India. Our study showed that compared with nonsmokers, smokers have an increased risk of hypermethylation in these genes. We found that 57.3% of the smokers samples showed methylation for GSTP1, 38% for p16, 18% for p14, and 32% for RASSF1A. Our population study allowed us to reveal the relationship between smoking and the subsequent appearance of an epigenetic change. Smoking speeds up the hypermethylation of these genes, which are thus unable to express, making the person more susceptible to the risk of lung and other solid carcinomas. Hypermethylation studies on DNA from two lung cancer cell lines (A549 and H460) were also done to compare the results, and the results are similar to samples of smokers. 1. Introduction Cancer is neither rare anywhere in the world, nor mainly confined to high-resource countries. The most commonly diagnosed cancers worldwide are lung followed by breast and colorectal cancers. Lung cancer is the most common cause of cancer-related mortality worldwide. About 1,80000 new cases are detected every year [1, 2]. Because of its high fatality (the ratio of mortality to incidence is 0.86) and the lack of variability in survival, in developed and developing countries, the highest and lowest mortality rates are estimated in the same regions, both in men and women. The large number of fatalities illustrates the lack of effective therapeutic alterations for a disease which is mostly diagnosed at an advanced stage [3]. There is a strong need for the development of biomarkers [4–6] that can spot this disease at an early stage which in turn would improve the survival rates. In comparison to mRNA, miRNA, and certain proteins, the use of genomic DNA methylation as biomarker has some novel attractions. Firstly, genomic DNA is highly stable, easy to extract, and secondly it can survive harsh conditions [7]. Genomic DNA has received special attention because of its potential application as a noninvasive, rapid, and sensitive tool which can lead to the development of clinically relevant biomarker for
References
[1]
J. Ferlay, H. R. Shin, F. Bray, D. Forman, C. Mathers, and D. M. Parkin, “Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008,” International Journal of Cancer, vol. 127, no. 12, pp. 2893–2917, 2010.
[2]
S. H. Landis, T. Murray, S. Bolden, and P. A. Wingo, “Cancer statistics, 1998,” Ca-A Cancer Journal for Clinicians, vol. 51, pp. 15–36, 2001, Erratum appears in Ca-A Cancer Journal for Clinicians, vol. 48, pp. 192, 1998.
[3]
G. M. Strauss, R. E. Gleason, and D. J. Sugarbaker, “Screening for lung cancer; another look: a different view,” Chest, vol. 113, no. 2, pp. 557–558, 1997.
[4]
D. Sidransky, “Emerging molecular markers of cancer,” Nature Reviews Cancer, vol. 2, no. 3, pp. 210–219, 2002.
[5]
P. W. Laird, “The power and the promise of DNA methylation markers,” Nature Reviews Cancer, vol. 3, no. 4, pp. 253–266, 2003.
[6]
S. A. Belinsky, K. J. Nikula, W. A. Palmisano et al., “Aberrant methylation of is an early event in lung cancer and a potential biomarker for early diagnosis,” Proceedings of the National Academy of Sciences of the United States of America, vol. 95, no. 20, pp. 11891–11896, 1998.
[7]
M. J. Duffy, R. Napieralski, J. W. M. Martens et al., “Methylated genes as new cancer biomarkers,” European Journal of Cancer, vol. 45, no. 3, pp. 335–346, 2009.
[8]
R. E. Board, L. Knight, A. Greystoke et al., “DNA methylation in circulating tumor DNA as a biomarker for cancer,” Biomark Insights, vol. 2, pp. 307–319, 2007.
[9]
E. Gormally, E. Caboux, P. Vineis, and P. Hainaut, “Circulating free DNA in plasma or serum as biomarker of carcinogenesis: practical aspects and biological significance,” Mutation Research, vol. 635, no. 2-3, pp. 105–117, 2007.
[10]
J. G. Herman and S. B. Baylin, “Gene silencing in cancer in association with promoter hypermethylation,” The New England Journal of Medicine, vol. 349, no. 21, pp. 2042–2054, 2003.
[11]
S. B. Baylin, J. G. Herman, J. R. Graff, P. M. Vertino, and J. P. Issa, “Alterations in DNA methylation: a fundamental aspect of neoplasia,” Advances in Cancer Research, vol. 72, pp. 141–196, 1997.
[12]
J. G. Herman, J. R. Graff, S. My?h?nen, B. D. Nelkin, and S. B. Baylin, “Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands,” Proceedings of the National Academy of Sciences of the United States of America, vol. 93, no. 18, pp. 9821–9826, 1996.
[13]
C. S. Morrow, K. H. Cowan, and M. E. Goldsmith, “Structure of the human genomic glutathione S-transferase-p1 gene,” Gene, vol. 75, no. 1, pp. 3–11, 1989.
[14]
T. R. Rebbeck, “Molecular epidemiology of the human glutathione S-transferase genotypes GSTM1 and GSTT1 in cancer susceptibility,” Cancer Epidemiology Biomarkers and Prevention, vol. 6, no. 9, pp. 733–743, 1997.
[15]
H. W. Lo, L. Stephenson, X. Cao, M. Milas, R. Pollock, and F. Ali-Osman, “Identification and functional characterization of the human Glutathione S-transferase P1 gene as a novel transcriptional target of the p53 tumor suppressor gene,” Molecular Cancer Research, vol. 6, no. 5, pp. 843–850, 2008.
[16]
R. A. Kratzke, T. M. Greatens, J. B. Rubins et al., “Rb and expression in resected non-small cell lung tumors,” Cancer Research, vol. 56, no. 15, pp. 3415–3420, 1996.
[17]
H. Tanaka, Y. Fujii, H. Hirabayashi et al., “Disruption of the RB pathway and cell-proliferative activity in non-small-cell lung cancers,” International Journal of Cancer, vol. 79, no. 2, pp. 111–115, 1998.
[18]
M. Serrano, “The tumor suppressor protein ,” Experimental Cell Research, vol. 237, no. 1, pp. 7–13, 1997.
[19]
M. Ruas and G. Peters, “The /CDKN2A tumor suppressor and its relatives,” Biochimica et Biophysica Acta, vol. 1378, no. 2, pp. F115–F177, 1998.
[20]
N. E. Sharpless and R. A. Depinho, “The INK4A/ARF locus and its two gene products,” Current Opinion in Genetics and Development, vol. 9, no. 1, pp. 22–30, 1999.
[21]
R. Dammann, C. Li, J. H. Yoon, P. L. Chin, S. Bates, and G. P. Pfeifer, “Epigenetic inactivation of a RAS association domain family protein from the lung tumour suppressor locus 3p21.3,” Nature Genetics, vol. 25, no. 3, pp. 315–319, 2000.
[22]
D. G. Burbee, E. Forgacs, S. Z?chbauer-Müller et al., “Epigenetic inactivation of RASSF1A in lung and breast cancers and malignant phenotype suppression,” Journal of the National Cancer Institute, vol. 93, no. 9, pp. 691–699, 2001.
[23]
S. J. Clark, A. Statham, C. Stirzaker, P. L. Molloy, and M. Frommer, “DNA methylation: bisulphite modification and analysis,” Nature Protocols, vol. 1, no. 5, pp. 2353–2364, 2006.
[24]
L. C. Li and R. Dahiya, “MethPrimer: designing primers for methylation PCRs,” Bioinformatics, vol. 18, no. 11, pp. 1427–1431, 2002.
[25]
F. Zhao, Z. Xuan, L. Liu, and M. Q. Zhang, “TRED: a Transcriptional Regulatory Element Database and a platform for in silico gene regulation studies,” Nucleic Acids Research, vol. 33, pp. D103–D107, 2005.
[26]
M. Esteller, “Epigenetic lesions causing genetic lesions in human cancer: promoter hypermethylation of DNA repair genes,” European Journal of Cancer, vol. 36, no. 18, pp. 2294–2300, 2000.
[27]
S. Z?chbauer-Müller, S. Lam, S. Toyooka et al., “Aberrant methylation of multiple genes in the upper aerodigestive tract epithelium of heavy smokers,” International Journal of Cancer, vol. 107, no. 4, pp. 612–616, 2003.
[28]
R. Peto, S. Darby, H. Deo, P. Silcocks, E. Whitley, and R. Doll, “Smoking, smoking cessation, and lung cancer in the UK since 1950: combination of national statistics with two case-control studies,” British Medical Journal, vol. 321, no. 7257, pp. 323–329, 2000.
[29]
W. A. Palmisano, K. K. Divine, G. Saccomanno et al., “Predicting lung cancer by detecting aberrant promoter methylation in sputum,” Cancer Research, vol. 60, no. 21, pp. 5954–5958, 2000.
[30]
V. Kulkarni and D. Saranath, “Concurrent hypermethylation of multiple regulatory genes in chewing tobacco associated oral squamous cell carcinomas and adjacent normal tissues,” Oral Oncology, vol. 40, no. 2, pp. 145–153, 2004.
[31]
J. S. Deep and S. Sidhu, “Methylation pattern of E-cadherin gene as epigenetic biomarker in lung cancer patients,” Research Journal of Biotechnology, vol. 3, no. 4, pp. 32–34, 2008.
[32]
J. S. Deep and S. Sidhu, “Study of methylation pattern of APC gene as epigenetic biomarker in lung cancer patients,” Research Journal of Biotechnology, vol. 3, pp. 435–437, 2008.
[33]
J. S. Deep, “DNA methylation: epigenetics to molecular diagnostics,” Research Journal of BioTechnology, vol. 4, no. 4, pp. 3–4, 2009.
[34]
S. Sidhu, J. S. Deep, R. C. Sobti, V. L. Sharma, and H. Thakur, “Methylation pattern of MGMT gene in relation to age, smoking, drinking and dietary habits as epigenetic biomarker in prostate cancer patients,” Genetic Engineering and Biotechnology Journal. In press.