Promoter hypermethylation of KLOTHO; an anti-senescence related gene in colorectal cancer patients of Kashmir valley

Document Type: Original article


1 Department of General and Minimal Access Surgery, SKIMS Srinagar, India

2 Department of Immunology and Molecular Medicine, SKIMS Srinagar, India


Hypermethylation of CpG islands located in the promoter regions of genes is a major event in the development of the majority of cancer types, due to the subsequent aberrant silencing of important tumor suppressor genes. KLOTHO; a novel gene associated primarily with suppressing senescence has been shown to contribute to tumorigenesis as a result of its impaired function. Recently the relevance of KLOTHO promoter hypermethylation in colorectal carcinoma in humans has been reported. We analyzed the promoter hypermethylation of KLOTHO gene in 50 histopathologically confirmed tumor and adjacent normal tissues of colorectal cancer patients. Methylation was assessed by bisulfite conversion of DNA followed by methylation specific-polymerase chain reaction. Methylation status was compared with gender, smoking status and histopathological parameters of patients. Promoter hypermethylation in KLOTHO gene was detected in 86% (43/50) of tumor tissues and 14% (7/50) of adjacent normal tissues. The methylation pattern differed significantly between tumor and adjacent normal tissues (P<0.0001). However, no association was found between promoter hypermethylation status and gender (P=0.68), smoking status (P=0.64) or other histopathological parameters (P>0.05) of colorectal cancer patients. We conclude that this novel tumor suppressor gene is epigenetically inactivated in colorectal cancer in our population paving way towards the potential of KLOTHO promoter hypermethylation as a predictor of the prognosis in colorectal cancer patients.


  1. Hamilton, SR, Aalton LA. Pathology and Genetics of Tumours of the Digestive System. IARC Press: Lyon; 2000.
  2. Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 2010; 127:2893-2917.
  3. Jemal A, Bray F, Center MM, FerlayJ, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin 2011;61:69-90.
  4. Mir MM, Dar NA, Gochhait S, Zargar SA, Ahangar AG, Bamezai RN. P53 mutation profile of squamous cell carcinoma of the esophagus in Kashmir (India): a high incidence area. Int J Cancer 2005;116:62-68.
  5. Murtaza I, Mushtaq D, Margoob MA, Dutt A, Wani NA, Ahmad I, Bhat ML. A study on P53 alterations in esophageal squamous cell carcinoma & their correlation to common dietary risk factors among population of the Kashmir valley. World J Gastroenterol 2006;12:4033-4037.
  6. Javid G, Zargar SA, Rather S, Khan A R, Khan B A,Yattoo GN, Shah A, Gulzar GM, Sodhi JS, Khan MA, Bashir AS. Incidence of CRC in Kashmir valley. Indian J Gastroenterol 2011;30:7-11.
  7. Matsumura Y, Aizawa H, Shiraki-Iida T, Nagai R, Kuro-o M, Nabeshima Y. Identification of the human klotho gene and its two transcripts encoding membrane and secreted klotho protein. Biochem Biophys Res Commun 1998;242:626-630.
  8. Wolf I, Levanon-Cohen S, Bose S, Ligumsky H, Sredni B, Kanety H, Kuro-o M, Karlan B, Kaufman B, Koeffler HP, Rubinek T. Klotho: a tumor suppressor and a modulator of the IGF-1 and FGF pathways in human breast cancer. Oncogene 2008; 27:7094-7105.
  9. Pan J, Zhong J, Gan LH, Chen SJ, Jin HC, Wang X, Wang LJ. Klotho, an anti-senescence related gene, is frequently inactivated through promoter hypermethylat-ion in colorectal cancer. Tumour Biol 2011;32:729-735.
  10. Wang L, Wang X, Wang X, Jie P, Lu H, Zhang S, Lin X, Lam EK, Cui Y, Yu J, Jin H. Klotho is silenced through promoter hypermethylation in gastric cancer. Am J Cancer Res 2011;1:111-119.
  11. Chen B, Ma X, Liu S, Zhao W, Wu J. Inhibition of lung cancer cells growth, motility and induction of apoptosis by Klotho, a novel secreted Wnt antagonist, in a dose-dependent manner. Cancer Biol Ther 2012;13:1221-1228.
  12. Klaus A, Birchmeier W. Wnt signalling and its impact on development and cancer. Nat Rev Cancer 2008;8:387-398.
  13. Liu H, Fergusson MM, Castilho RM, Liu J, Cao L, Chen J, Malide D, Rovira II, Schimel D, Kuo CJ, Gutkind JS, Hwang PM, Finkel T. Augmented Wnt signaling in a mammalian model of accelerated aging. Science 2007;317:803-806.
  14. Lee J, Jeong DJ, Kim J, Lee S, Park JH, Chang B, Jung SI, Yi L, Han Y, Yang Y, Kim KI, Lim JS, Yang I, Jeon S, Bae DH, Kim CJ, Lee MS. The anti-aging gene KLOTHO is a novel target for epigenetic silencing in human cervical carcinoma. Mol Cancer 2010;9:109.
  15. Gan LH, Pan J, Chen SJ, Zhong J, Wang LJ. DNA methylation of ZIC1 and KLOTHO gene promoters in colorectal carcinomas and its clinicopathological significance. Zhejiang Da Xue Xue Bao Yi Xue Ban 2011;40:309-314.
  16. Xie B, Zhou J, Shu G, Liu DC, Zhou J, Chen J, Yuan L. Restoration of klotho gene expression induces apoptosis and autophagy in gastric cancer cells: tumor suppressive role of Klotho in gastric cancer. Cancer Cell Int 2013;13:18.
  17. Wani HA, Bhat AA, Mattoo AA, Khan H, Amin S, Bhat SA, Naikoo NA, Rasheed T, Masood A, Majid S. Distribution of P16 promoter hypermethylation in male/female colorectal cancer patients of Kashmir Valley. Int J Eng Sci Invent 2013;2:11-17.
  18. Bird A. Perceptions of epigenetics. Nature 2007;447:396-398.
  19. Esteller M. Epigenetics in cancer. N Engl J Med 2008;358:1148-1159.
  20. Hoeijmakers JH. DNA damage, aging and cancer. N Engl J Med 2009;361:1475-1485.
  21. Collado M, Serrano M. Senescence in tumours: evidence from mice and humans. Nat Rev Cancer 2010;10:51–57.
  22. Kuroo M, Matsumura Y, Aizawa H, Kawaguchi H, Suga T, Utsugi T, Ohyama Y, Kurabayashi M, Kaname T, Kume E, Iwasaki H, Iida A, Shiraki-Iida T, Nishikawa S, Nagai R, Nabeshima YI. Mutation of the mouse Klotho gene leads to a syndrome resembling ageing. Nature1997; 390:45–51.
  23. Kurosu H, Yamamoto M, Clark JD, Pastor JV, Nandi A, Gurnani P, McGuinness OP, Chikuda H, Yamaguchi M, Kawaguchi H, Shimomura I, Takayama Y, Herz J, Kahn CR, Rosenblatt KP, Kuroo M. Suppression of aging in mice by the hormone Klotho. Science 2005;309:1829-1833.
  24. Chan AT, Giovannucci EL. Primary prevention of colorectal cancer. Gastroenterology 2010;138:2029-2043.
  25. Wilkins HR, Doucet K, Duke V, Morra A, Johnson N. Estrogen prevents sustained COLO-205 human colon cancer cell growth by inducing apoptosis, decreasing c-myb protein, and decreasing transcription of the anti-apoptotic protein bcl-2. Tumour Biol 2010;31:16-22.
  26. Martineti V, Silvestri S, Tonelli F, Brandi ML. Control of colon cancer development and progression by selected estrogen receptor modulators. Expert Rev Endocrinol Metab 2008;3:503-511. 
  27. Grady WM. Genetic testing for high-risk colon cancer patients. Gastroenterology 2003;124:1574-1594.
  28. Sondergaard JO, Bulow S, Lynge E. Cancer incidence among parents of patients with colorectal cancer. Int J Cancer 1991;47:202-206.
  29. Fuchs CS, Giovannucci EL, Colditz GA, Hunter DJ, Speizer FE, Willett WC. A prospective study of family history and the risk of colorectal cancer. N Engl J Med 1994;331:1669-1674.
  30. Botteri E, Iodice S, Bagnardi V, Raimondi S, Lowenfels AB, Maisonneuve P. Smoking and colorectal cancer: a meta-analysis. JAMA 2008;300:2765-2778.