Loss of p15INK4b Expression in Colorectal Cancer is Linked to Ethnic Origin

  • Abdel-Rahman, Wael Mohamed (College of Health Sciences, University of Sharjah, and Sharjah Institute for Medical Research) ;
  • Nieminen, Taina Tuulikki (Department of Medical Genetics, University of Helsinki) ;
  • Shoman, Soheir (Department of Pathology, National Cancer Institute, Cairo University) ;
  • Eissa, Saad (Department of Pathology, National Cancer Institute, Cairo University) ;
  • Peltomaki, Paivi (Department of Medical Genetics, University of Helsinki)
  • Published : 2014.03.01


Colorectal cancers remain to be a common cause of cancer-related death. Early-onset cases as well as those of various ethnic origins have aggressive clinical features, the basis of which requires further exploration. The aim of this work was to examine the expression patterns of $p15^{INK4b}$ and SMAD4 in colorectal carcinoma of different ethnic origins. Fifty-five sporadic colorectal carcinoma of Egyptian origin, 25 of which were early onset, and 54 cancers of Finnish origin were immunohistochemically stained with antibodies against $p15^{INK4b}$ and SMAD4 proteins. Data were compared to the methylation status of the $p15^{INK4b}$ gene promotor. $p15^{INK4b}$ was totally lost or deficient (lost in ${\geq}50%$ of tumor cell) in 47/55 (85%) tumors of Egyptian origin as compared to 6/50 (12%) tumors of Finnish origin (p=7e-15). In the Egyptian cases with $p15^{INK4b}$ loss and available $p15^{INK4b}$ promotor methylation status, 89% of cases which lost $p15^{INK4b}$ expression were associated with $p15^{INK4b}$ gene promotor hypermethylation. SMAD4 was lost or deficient in 25/54 (46%) tumors of Egyptian origin and 28/48 (58%) tumors of Finnish origin. 22/54 (41%) Egyptian tumors showed combined loss/deficiency of both $p15^{INK4b}$ and SMAD4, while $p15^{INK4b}$ was selectively lost/deficient with positive SMAD4 expression in 24/54 (44%) tumors. Loss of $p15^{INK4b}$ was associated with older age at presentation (>50 years) in the Egyptian tumors (p=0.04). These data show for the first time that $p15^{INK4b}$ loss of expression marks a subset of colorectal cancers and ethnic origin may play a role in this selection. In a substantial number of cases, the loss was independent of SMAD4 but rather associated with $p15^{INK4b}$ gene promotor hypermethylation and old age which could be related to different environmental exposures.


  1. Simon M, Park TW, Koster G, et al (2001). Alterations of INK4a(p16-p14ARF)/INK4b(p15) expression and telomerase activation in meningioma progression. J Neurooncol, 55, 149-58.
  2. Senut MC, Sen A, Cingolani P, et al (2014). Lead exposure disrupts global dna methylation in human embryonic stem cells and alters their neuronal differentiation. Toxicol Sci, [Epub ahead of print].
  3. Seoane J, Pouponnot C, Staller P, et al (2001). TGFbeta influences MYC, Miz-1 and Smad to control the CDK inhibitor $p15^{INK4b}$. Nat Cell Biol, 3, 400-8.
  4. Teofili L, Morosetti R, Martini M, et al (2000). Expression of cyclin-dependent kinase inhibitor $p15^{(INK4B)}$ during normal and leukemic myeloid differentiation. Exp Hematol, 28, 519-26.
  5. Warner BJ, Blain SW, Seoane J, et al (1999). MYC downregulation by transforming growth factor beta required for activation of the p15(Ink4b) G(1) arrest pathway. Mol Cell Biol, 19, 5913-22.
  6. Xu XL, Yu J, Zhang HY, et al (2004). Methylation profile of the promoter CpG islands of 31 genes that may contribute to colorectal carcinogenesis. World J Gastroenterol, 10, 3441-54.
  7. Zekri Ael R, Nassar AA, El-Din El-Rouby MN, et al (2013). Disease progression from chronic hepatitis C to cirrhosis and hepatocellular carcinoma is associated with increasing DNA promoter methylation. Asian Pac J Cancer Prev, 14, 6721-6.
  8. Zhu QC, Gao RY, Wu W, et al (2013). Epithelial-mesenchymal transition and its role in the pathogenesis of colorectal cancer. Asian Pac J Cancer Prev, 14, 2689-98.
  9. O'Hagan HM (2013). Chromatin modifications during repair of environmental exposure-induced DNA damage: A potential mechanism for stable epigenetic alterations. Environ Mol Mutagen. 55, 278-91
  10. Moad AI, Lan TM, Kaur G, et al (2009). Immunohistochemical determination of the P15 protein expression in cutaneous squamous cell carcinoma. J Cutan Pathol, 36, 183-9.
  11. Nakamura H, Makino K, Kuratsu J (2011). Molecular and clinical analysis of glioblastoma with an oligodendroglial component (GBMO). Brain Tumor Pathol, 28, 185-90.
  12. Nassiri M, Kooshyar MM, Roudbar Z, et al (2013). Genes and SNPs associated with non-hereditary and hereditary colorectal cancer. Asian Pac J Cancer Prev, 14, 5609-14.
  13. Nieminen TT, Shoman S, Eissa S, et al (2012). Distinct genetic and epigenetic signatures of colorectal cancers according to ethnic origin. Cancer Epidemiol Biomarkers Prev, 21, 202-11.
  14. Oda Y, Yamamoto H, Takahira T, et al (2005). Frequent alteration of p16(INK4a)/p14(ARF) and p53 pathways in the round cell component of myxoid/round cell liposarcoma: p53 gene alterations and reduced p14(ARF) expression both correlate with poor prognosis. J Pathol, 207, 410-21.
  15. Paun BC, Kukuruga D, Jin Z, et al (2010). Relation between normal rectal methylation, smoking status, and the presence or absence of colorectal adenomas. Cancer, 116, 4495-501.
  16. Royce SG, Alsop K, Haydon A, et al (2010). The role of SMAD4 in early-onset colorectal cancer. Colorectal Dis, 12, 213-9.
  17. Sakellariou S, Liakakos T, Ghiconti I, et al (2008). Immunohistochemical expression of P15 (INK4B) and SMAD4 in advanced gastric cancer. Anticancer Res, 28, 1079-83.
  18. Holm R, Forsund M, Nguyen MT, et al (2013). Expression of p15INK(4)b and p57KIP(2) and relationship with clinicopathological features and prognosis in patients with vulvar squamous cell carcinoma. PLoS One, 8, 61273.
  19. elinsky SA, Klinge DM, Liechty KC, et al (2004). Plutonium targets the p16 gene for inactivation by promoter hypermethylation in human lung adenocarcinoma. Carcinogenesis, 25, 1063-7.
  20. Fraga MF, Agrelo R and Esteller M (2007). Cross-talk between aging and cancer: the epigenetic language. Ann N Y Acad Sci, 1100, 60-74.
  21. Herman JG, Civin CI, Issa JPJ, et al (1997). Distinct patterns of inactivation of $p15^{(INK4B)}$ and $p16^{(INK4A)}$ characterize the major types of hematological malignancies. Cancer Research, 57, 837-41.
  22. Ishiguro A, Takahata T, Saito M, et al (2006). Influence of methylated p15 and p16 genes on clinicopathological features in colorectal cancer. J Gastroenterol Hepatol, 21, 1334-9.
  23. Joensuu EI, Abdel-Rahman WM, Ollikainen M, et al (2008). Epigenetic signatures of familial cancer are characteristic of tumor type and family category. Cancer Res, 68, 4597-605.
  24. Kim WY and Sharpless NE (2006). The regulation of INK4/ ARF in cancer and aging. Cell, 127, 265-75.
  25. Lampropoulos P, Zizi-Sermpetzoglou A, Rizos S, et al (2012). TGF-beta signalling in colon carcinogenesis. Cancer Lett, 314, 1-7.
  26. Lu SL, Akiyama Y, Nagasaki H, et al (1995). Mutations of the transforming growth factor-beta type II receptor gene and genomic instability in hereditary nonpolyposis colorectal cancer. Biochem Biophys Res Commun, 216, 452-7.
  27. Marsit CJ, Karagas MR, Danaee H, et al (2006). Carcinogen exposure and gene promoter hypermethylation in bladder cancer. Carcinogenesis, 27, 112-6.
  28. Bodoor K, Haddad Y, Alkhateeb A, et al (2014). DNA Hypermethylation of Cell Cycle (p15 and p16) and Apoptotic (p14, p53, DAPK and TMS1) Genes in Peripheral Blood of Leukemia Patients. Asian Pac J Cancer Prev, 15, 75-84.
  29. Cheng YW, Shawber C, Notterman D, et al (2006). Multiplexed profiling of candidate genes for CpG island methylation status using a flexible PCR/LDR/Universal Array assay. Genome Res, 16, 282-9.
  30. Coppede F, Migheli F, Lopomo A, et al (2014). Gene promoter methylation in colorectal cancer and healthy adjacent mucosa specimens: Correlation with physiological and pathological characteristics, and with biomarkers of onecarbon metabolism. Epigenetics, 9 [Epub ahead of print].
  31. Endo M, Kobayashi C, Setsu N, et al (2011). Prognostic significance of p14ARF, $p15^{INK4b}$, and p16INK4a inactivation in malignant peripheral nerve sheath tumors. Clin Cancer Res, 17, 3771-82.
  32. Esteller M, Corn PG, Baylin SB, et al (2001). A gene hypermethylation profile of human cancer. Cancer Res, 61, 3225-9.
  33. Ahn BK, Jang SH, Paik SS, et al (2011). Smad4 may help to identify a subset of colorectal cancer patients with early recurrence after curative therapy. Hepatogastroenterology, 58, 1933-6.

Cited by

  1. The Role of Stem Cell DNA Methylation in Colorectal Carcinogenesis vol.12, pp.5, 2016,