Skip to main content

Advertisement

SpringerLink
Log in

Aberrant p16INK4a methylation is a frequent event in colorectal cancers: prognostic value and relation to mRNA expression and immunoreactivity

  • Original Paper
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Purpose

Aberrant p16INK4a promoter methylation is common in colorectal cancer (CRC), but its clinicopathological significance remains controversial. The present study was therefore conducted to analyze p16INK4a methylation and its relationship to clinicopathological features, mRNA levels and immunoreactivity in a series of lesions.

Methods

p16INK4a methylation was assessed for normal mucosa (n = 30) and CRC samples (n = 212) by methylation-specific real-time quantitative PCR, and p16INK4a expression by immunostaining in formalin-fixed paraffin-embedded specimens. In addition, fresh DNA (n = 61) was analyzed for relationships to p16INK4a mRNA by reverse-transcription PCR.

Results

The p16INK4a methylation index of normal mucosa samples ranged from 0 to 2% (mean, 0.23%; median, 0.02%), while the values for tumor samples varied widely from 0 to 100% (mean, 25.7%; median, 7.1%), the difference being statistically significant (P < 0.001). Of 151 paraffin-embedded CRC tissue samples, 51 (34%), 54 (36%), and 46 (30%) were classified as low, intermediate, and high for aberrant methylation of p16INK4a. High p16INK4a methylation was significantly associated with large tumor size (P = 0.025). Patients with higher methylation further showed more frequent recurrence as compared with the low-methylation group, and shortened cancer-related survival (Hazard ratio [HR], 3.379; P < 0.001) and recurrence-free survival (HR, 3.962; P < 0.001 on multivariate analysis). A significant inverse relationship was apparent between the p16INK4a methylation and immunoreactivity (P = 0.017). A similar tendency was also observed for the methylation status and the mRNA level (P = 0.195).

Conclusions

We conclude that p16INK4a methylation results in transcriptional silencing and defines a group of CRCs with a poor prognosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Ahuja N, Mohan AL, Li Q, Stolker JM, Herman JG, Hamilton SR, Baylin SB, Issa J-PJ (1997) Association between CpG island methylation and microsatellite instability in colorectal cancer. Cancer Res 57:3370–3374

    CAS  PubMed  Google Scholar 

  • Baylin SB, Herman JG, Graff JR, Vertino PM, Issa J-P (1998) Alterations in DNA methylation: a fundamental aspect of neoplasia. Adv Cancer Res 72:141–196

    Article  CAS  PubMed  Google Scholar 

  • Burri N, Shaw P, Bouzourene H, Sordat I, Sordat B, Gillet M, Schorderet D, Bosman FT, Chaubert P (2001) Methylation silencing and mutations of the p14 ARF and p16 INK4a genes in colon cancer. Lab Invest 81:217–229

    CAS  PubMed  Google Scholar 

  • Dai CY, Furth EE, Mick R, Koh J, Takayama T, Niitsu Y, Enders GH (2000) p16INK4a expression begins early in human colon neoplasia and correlates inversely with markers of cell proliferation. Gastroenterology 119:929–942

    Article  CAS  PubMed  Google Scholar 

  • Derks S, Postma C, Moerkerk PTM, van den Bosch SM, Carvalho B, Hermsen MAJA, Giaretti W, Herman JG, Weijenberg MP, de Bruïne AP, Meijer GA, van Engeland M (2006) Promoter methylation precedes chromosomal alterations in colorectal development. Cellular Oncol 28:247–257

    CAS  Google Scholar 

  • Esteller M, Tortola S, Toyota M, Capella G, Peinado MA, Baylin SB, Herman JG (2000) Hypermethylation-associated inactivation of p14 ARF is independent of p16 INK4a methylation and p53 mutational status. Cancer Res 60:129–133

    CAS  PubMed  Google Scholar 

  • Esteller M, González S, Risques RA, Marcuello E, Mangues R, Germà JR, Herman JG, Capellà G, Peinado MA (2001) K-ras and p16 aberrations confer poor prognosis in human colorectal cancer. J Clin Oncol 19:299–304

    CAS  PubMed  Google Scholar 

  • Goto T, Mizukami H, Shirahata A, Sakata M, Saito M, Ishibashi K, Kigawa G, Nemoto H, Sanada Y, Hibi K (2009) Aberrant methylation of the p16 gene is frequently detected in advanced colorectal cancer. Anticancer Res 29:275–278

    CAS  PubMed  Google Scholar 

  • Guan RJ, Fu Y, Holt PR, Pardee AB (1999) Association of k-ras with p16 methylation in human colon cancer. Gastroenterology 116:1063–1071

    Article  CAS  PubMed  Google Scholar 

  • Hawkins N, Norrie M, Cheong K, Mokany E, Ku S-L, Meagher A, O’Connor T, Ward R (2002) CpG island methylation in sporadic colorectal cancers and its relationship to microsatellite instability. Gastroenterology 122:1376–1387

    Article  CAS  PubMed  Google Scholar 

  • Herman JG, Merlo A, Mao L, Lapidus RG, Issa J-PJ, Davidson NE, Sidransky D, Baylin SB (1995) Inactivation of the CDK2/p16/MTS1 gene is frequently associated with aberrant DNA methylation in all common human cancers. Cancer Res 55:4525–4530

    CAS  PubMed  Google Scholar 

  • Herman JG, Graff JR, Myöhänen S, Nelkin BD, Baylin SB (1996) Methylation-specific PCR: A novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA 93:9821–9826

    Article  CAS  PubMed  Google Scholar 

  • Hibi K, Nakayama H, Koike M, Kasai Y, Ito K, Akiyama S, Nakao A (2002) Colorectal cancers with both p16 and p14 methylation show invasive characteristics. Jpn J Cancer Res 93:883–887

    CAS  PubMed  Google Scholar 

  • Iacopetta B, Grieu F, Li W, Ruszkiewicz A, Caruso M, Moore J, Watanabe G, Kawakami K (2006) APC gene methylation is inversely correlated with features of the CpG island methylator phenotype in colorectal cancer. Int J Cancer 119:2272–2278

    Article  CAS  PubMed  Google Scholar 

  • International Union Against Cancer (UICC) (1997) TNM classification of the tumours of the colon and rectum. In: Sobin LH, Wittekind Ch (eds) TNM Classification of Malignant Tumours, 5th edn. Wiley-Liss Inc., New York, pp 59–62

  • Ishiguro A, Takahata T, Saito M, Yoshiya G, Tamura Y, Sasaki M, Munakata A (2006) Influence of methylated p15 INK4b and p16 INK4a genes on clinicopathological features in colorectal cancer. J Gastroenterol Hepatol 21:1334–1339

    Article  CAS  PubMed  Google Scholar 

  • Kamb A, Gruis NA, Weaver-Feldhaus J, Liu Q, Harshman K, Tavtigian SV, Stockert E, Day IIIRS, Johnson BE, Skolnick MH (1994) A cell cycle regulator potentially involved in genesis of many tumor types. Science 264:436–440

    Article  CAS  PubMed  Google Scholar 

  • Kanai Y, Ushijima S, Kondo Y, Nakanishi Y, Hirohashi S (2001) DNA methyltransferase expression and DNA methylation of CpG islands and peri-centromeric satellite regions in human colorectal and stomach cancers. Int J Cancer 91:205–212

    Article  CAS  PubMed  Google Scholar 

  • Kim BN, Yamamoto H, Ikeda K, Damdinsuren B, Sugita Y, Ngan CY, Fujie Y, Ogawa M, Hata T, Ikeda M, Ohue M, Sekimoto M, Monden T, Matsuura N, Monden M (2005) Methylation and expression of p16INK4 tumor suppressor gene in primary colorectal cancer tissues. Int J Oncol 26:1217–1226

    CAS  PubMed  Google Scholar 

  • Lee M, Han WS, Kim OK, Sung SH, Cho MS, Lee SN, Koo H (2006) Prognostic value of p16 INK4a and p14 ARF gene hypermethylation in human colon cancer. Pathol Res Pract 202:415–424

    Article  CAS  PubMed  Google Scholar 

  • Liang J-T, Chang K-J, Chen J-C, Lee C–C, Cheng Y-M, Hsu H-C, Wu M-S, Wang S-M, Lin J-T, Cheng A-L (1999) Hypermethylation of the p16 gene in sporadic T3N0M0 stage colorectal cancers: association with DNA replication error and shorter survival. Oncology 57:149–156

    Article  CAS  PubMed  Google Scholar 

  • Maeda K, Kawakami K, Ishida Y, Ishiguro K, Omura K, Watanabe G (2003) Hypermethylation of the CDKN2A gene in colorectal cancer is associated with shorter survival. Oncol Rep 10:935–938

    CAS  PubMed  Google Scholar 

  • Nobori T, Miura K, Wu DJ, Lois A, Takabayashi K, Carson DA (1994) Deletions of the cyclin-dependent kinase-4 inhibitor gene in multiple human cancers. Nature 368:753–756

    Article  CAS  PubMed  Google Scholar 

  • Norrie MWA, Hawkins NJ, Todd AV, Meagher AP, O’Connor TW, Ward RL (2003) Inactivation of p16 INK4a by CpG hypermethylation is not a frequent event in colorectal cancer. J Surg Oncol 84:143–150

    Article  CAS  PubMed  Google Scholar 

  • Ogino S, Odze RD, Kawasaki T, Brahmandam M, Kirkner GJ, Laird PW, Loda M, Fuchs CS (2006) Correlation of pathologic features with CpG island methylator phenotype (CIMP) by quantitative DNA methylation analysis in colorectal carcinoma. Am J Surg Pathol 30:1175–1183

    PubMed  Google Scholar 

  • Prall F, Ostwald C, Weirich V, Nizze H (2006) p16INK4a promoter methylation and 9q21 allelic loss in colorectal carcinomas: relation with immunohistochemical p16INK4a expression and with tumor budding. Hum Pathol 37:578–585

    Article  CAS  PubMed  Google Scholar 

  • Sanz-Casla MT, Maestro ML, Vidaurreta M, Maestro C, Arroyo M, Cerdán J (2005) p16 gene methylation in colorectal tumors: correlation with clinicopathological features and prognostic value. Dig Dis 23:151–155

    Article  CAS  PubMed  Google Scholar 

  • Schneider-Stock R, Boltze C, Peters B, Höpfner T, Meyer F, Lippert H, Roessner A (2003) Differences in loss of p16INK4 protein expression by promoter methylation between left- and right-sided primary colorectal carcinomas. Int J Oncol 23:1009–1013

    CAS  PubMed  Google Scholar 

  • Serrano M, Gómez-Lahoz E, DePinho RA, Beach D, Bar-Sagi D (1995) Inhibition of ras-induced proliferation and cellular transformation by p16INK4. Science 267:249–252

    Article  CAS  PubMed  Google Scholar 

  • Shannon BA, Iacopetta BJ (2001) Methylation of the hMLH1, p16, and MDR1 genes in colorectal carcinoma: associations with clinicopathological features. Cancer Lett 167:91–97

    Article  CAS  PubMed  Google Scholar 

  • Shen L, Catalano PJ, Benson AB III, O’Dwyer P, Hamilton SR, Issa J-PJ (2007) Association between DNA methylation and shortened survival in patients with advanced colorectal cancer treated with 5-fluorouracil-based chemotherapy. Clin Cancer Res 13:6093–6098

    Article  CAS  PubMed  Google Scholar 

  • Toyota M, Ahuja N, Ohe-Toyota M, Herman JG, Baylin SB, Issa J-PJ (1999) CpG island methylator phenotype in colorectal cancer. Proc Natl Acad Sci USA 96:8681–8686

    Article  CAS  PubMed  Google Scholar 

  • Van Rijnsoever M, Grieu F, Elsaleh H, Joseph D, Iacopetta B (2002) Characterisation of colorectal cancers showing hypermethylation at multiple CpG islands. Gut 51:797–802

    Article  PubMed  Google Scholar 

  • Van Rijnsoever M, Elsaleh H, Joseph D, McCaul K, Iacopetta B (2003) CpG island methylator phenotype is an independent predictor of survival benefit from 5-fluorouracil in stage III colorectal cancer. Clin Cancer Res 9:2898–2903

    PubMed  Google Scholar 

  • Ward RL, Cheong K, Ku S-L, Meagher A, O’Connor T, Hawkins NJ (2003) Adverse prognostic effect of methylation in colorectal cancer is reversed by microsatellite instability. J Clin Oncol 21:3729–3736

    Article  CAS  PubMed  Google Scholar 

  • Weinberg RA (1995) The retinoblastoma protein and cell cycle control. Cell 81:323–330

    Article  CAS  PubMed  Google Scholar 

  • Wiencke JK, Zheng S, Lafuente A, Lafuente MJ, Grudzen C, Wrensch MR, Miike R, Ballesta A, Trias M (1999) Aberrant methylation of p16 INK4a in anatomic and gender-specific subtypes of sporadic colorectal cancer. Cancer Epidemiol Biomark Prev 8:501–506

    CAS  Google Scholar 

  • Yi J, Wang Z-W, Cang H, Chen Y-Y, Zhao R, Yu B-M, Tang X-M (2001) p16 gene methylation in colorectal cancers associated with Duke’s staging. World J Gastroenterol 7:722–725

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a Grants-in-Aid from the Japan Society for the Promotion of Science (#21590394) to H. Mitomi. We thank Y. Oya, J. Kubo and N. Anpo, Pathology Division, National Hospital Organization Sagamihara Hospital, for their expert technical assistance.

Conflict of Interest Statement

None

Author information

Authors and Affiliations

  1. Department of Human Pathology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo, 113-8421, Japan

    Hiroyuki Mitomi, Tsuyoshi Saito, Takashi Matsuoka & Takashi Yao

  2. Department of Pathomechanisms, Clinical Research Center, National Hospital Organization, Sagamihara Hospital, Sagamihara, Japan

    Naoshi Fukui & Nobuho Tanaka

  3. Department of Surgery, National Hospital Organization, Sagamihara Hospital, Sagamihara, Japan

    Hideki Kanazawa

Authors
  1. Hiroyuki Mitomi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Naoshi Fukui
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nobuho Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hideki Kanazawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tsuyoshi Saito
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Takashi Matsuoka
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Takashi Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hiroyuki Mitomi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mitomi, H., Fukui, N., Tanaka, N. et al. Aberrant p16INK4a methylation is a frequent event in colorectal cancers: prognostic value and relation to mRNA expression and immunoreactivity. J Cancer Res Clin Oncol 136, 323–331 (2010). https://doi.org/10.1007/s00432-009-0688-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00432-009-0688-z

Keywords

Search

Navigation