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Aberrant methylation in promoter region induced silencing of P16/NK4a in colorectal cancer in Iranian patients

Jamshid Mehrzad, Mohammad Hashemi, Mahmodreza Jamshidi, Alireza motavalizadekakhki, Jafar Saeidi, Mahdieh Mohammaditabr, Arezoo Sadat Khafi, Behzad Taheri Moghaddam

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J. Bio. Env. Sci.4(2), 94-103, February 2014


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Aberrant methylation in promoter regions of genes might lead to change gene functions and result in cancer. Hence, biomarker identification for aberrant methylated genes would be very useful for early diagnosis, prognosis, and therapeutic treatment of colorectal cancer (CRC). The aim of the present study was to detection correlation between methylation status in promoter region of P16/NK4a with P16/NK4a expression level, CRC occurence and with demographic and clinocopathological characteristics of CRC. Methylation status in promoter region of P16/NK4a was assayed by methylation-specific polymerase chain reaction (MS-PCR) and P16/NK4a gene expression was performed by real time quantitative PCR (qPCR). RNA from embedded paraffin sections of colorectal tissue (in 70 sporadic colorectal tumors as well as adjoining and normal tissue specimens) was reverse transcribed, quantified and analyzed by Q-PCR. Aberrant promoter methylation of p16 gene was detected in 27 (38.6%) tumor samples and in 8 (11.4%) adjacent normal tissues. Thus, aberrant promoter methylation of p16 is significantly correlated with CRC occurance. Aberrant promoter methylation was found significantly associated with tumor stage II (P=0.000), but not with other clinocopathological and demographic characteristics. P16/INK4a expression level in tumor tissues was 8.6-fold more than normal adjacent tissues. In conclusion, this study has identified aberrant promoter methylation of P16/INK4a was significantly correlated with CRC, because aberrant promoter methylation effects on P16/INK4a expression. Our approaches revealed P16/INK4a can be as a potential biomarker for CRC as diagnostic, prognostic and therapeutic targets in the future.


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Aberrant methylation in promoter region induced silencing of P16/NK4a in colorectal cancer in Iranian patients

An C, Choi IS, Yao JC, Worah S, Xie K, Mansfield PF. 2005. Prognostic significance of CpG island methylator phenotype and microsatellite instability in gastric carcinoma. Clinical Cancer Research 11, 656-663.

Ashktorab H, Smoot DT, Carethers JM, Rahmanian M, Kittles R, Vosganian G, Doura M, Nidhiry E, Naab T, Momen B. 2003. Highmincidence of microsatellite instability in colorectal cancer from African Americans. Clinical Cancer Research 9, 1112-1117.

Baylin SB. 2005. DNA methylation and gene silencing in cancer. Nature Clinical Practice Oncology 2, 104-111.

Baylin SB, Ohm JE. 2006. Epigenetic gene silencing in cancer – a mechanism for early oncogenic pathway addiction? Nature Reviews Cancer 6, 107-116.

Caradec J, Sirab N, Revaud D, Keumeugni C, Loric S. 2010. Is GAPDH a relevant housekeeping gene for normalization in colorectal cancer experiments? British Journal of Cancer 103, 1475 – 1476.

Changmei G, Jiachun L, Tianpen C, Cheng L, Hao S, Wenmao X, Xueli Y, Xiaobo Y, Yangxin H, Meixia L. 2013. Association between MGMT Promoter Methylation and Non-Small Cell Lung Cancer: A Meta-Analysis. PLOS ONE 8 (9), 72-633.

Edge SB, Byrd DR, Compton CC, Fritz AG, Greene FL. 2010. Cancer Staging Handbook: From the AJCC Cancer Staging Manual.: Springer Publishing Company, 173–206

El-Naggar AK, Lai S, Clayman G, Lee JK, Luna MA, Goepfert H, Batsakis JG. 1997. Methylation, a major mechanism of p16/CDKN2 gene inactivation in head and neck squamous carcinoma. American Journal of Pathology 151, 1767-1774.

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. Journal of Clinical Oncology 19, 299-304.

Flood DM, Weiss NS, Cook LS. 2000. Colorectal cancer incidence in Asian migrants to the United States and their descendants. Cancer Causes Control 11(5), 403–411.

Fountain JW, Karayiogou M, Ernstoff MS, Kirkwood JM, Viock DR,Titus-Emstoff,L, Bouchard B, Vijayasaradh S, Houghton AN, Lahti,J. 1992. Homozygous deletions within human chromosome band 9p21 in melanoma. of the National Academy Sciences USA 89, 1055-10561.

Grulich AE, McCredie M, Coates M. 1995. Cancer incidence in Asian migrants to New South Wales. Australia. British Journal of Cancer 71(2), 400–408.

Jemal A, Bray F, Center MM. 2011. Global cancer statistics. Cancer Journal for Clinicians 61(2), 69– 90.

Herman JG, Graff JR, Myohanen S. 1996. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proceedings of the National Academy Sciences USA 93, 98-216.

Ivanova TA, Golovina DA, Zavalishina LE, Volgareva GM, Katargin AL, Andreeva YY, Frank GA, Kisseljov FL, Kisseljova NP. 2007. Up-regulation of expression and lack of 5′ CpG island hypermethylation of p16 INK4a in HPV-positive cervical carcinomas. BMC Cancer 7, 47.

Jundong G, Yanjun W, Siwei Z. Feng H, Hui Z, Hongrui X, Jiacong Y,Linlin S, Weiqiang W, Jun C, Qinghua Z. 2013. Association between P16INK4a Promoter Methylation and Non-Small Cell Lung Cancer: A Meta-Analysis. PLOS ONE 4, 60-107.

Khor GH. 2009. Chromosome 17 Aberration of Oral Squamous Cell Carcinoma in Malaysia. Global Journal of Health Science 1, 150-156.

Khor GH, Hassan MI, Siar CH. 2011. p53 Expression as a marker of microinvasion in oral squamous cell carcinoma. Asian Pacific Journal of Cancer Prevention 12, 1017-1022.

Lee J, Demissie K, Lu SE, Rhoads GG. 2007. Cancer incidence among Korean-American immigrants in the United States and native Koreans in South Korea. Cancer Control 14(1), 78–85.

Li Z, Fang ZY, Ding Y, Yao WT, Yang Y. 2012. Amplifications of NCOA3 gene in colorectal cancers in a Chinese population. World Journal of Gastroenterology 18, 855–860.

Liang JT, Chang KJ, Chen JC, Lee CC, Cheng YM, Hsu HC, Wu MS, Wang SM, Lin JT, Cheng AL. 1999. Hypermethylation of the p16 gene in sporadic T3N0M0 stage colorectal cancers: association with DNA replication error and shorter survival. Oncology 57, 149-156.

Matsuda Y, Ichida T, Matsuzawa J, Sugimura K, Asakura H. 1999. p16(INK4) is inactivated by extensive CpG methylation in human hepatocellular carcinoma. Gastroenterology 116, 394-400.

Miyamoto K, Ushijima T. 2005. Diagnostic and Therapeutic Applications of Epigenetics. Japanese Journal of Clinical Oncology 35, 293-301.

Radhakrishnan R, Kabekkodu S, Satyamoorthy K. 2011. DNA hypermethylation as an epigenetic mark for oral cancer diagnosis. Journal of Oral Pathology & Medicine 40, 665-676.

Mulero-Navarro S, Esteller M. 2008. Epigenetic biomarkers for human cancer: The time is now. Critical Reviews in Oncology/Hematology 68, 1-11.

Nakahara Y, Shintani S, Mihara M, Ueyama Y, Matsumura T. 2001. High frequency of homozygous deletion and methylation of p16(INK4A) gene in oral squamous cell carcinomas. Cancer Letter 163, 221-228.

Nakamura M, Sugita K, Inukai T, Goi K, Iijima K, Tezuka T, Kojika S, Shiraishi K, Miyamoto N, Karakida N. 1999. p16/MTS1/INK4A gene is frequently inactivated by hypermethylation in childhood acute lymphoblastic leukemia with 11q23 translocation. Leukemia 13, 884-890.

Nielsen NH, Roos G, Emdin SO, Landberg G. 2001. Methylation of the p16(Ink4a) tumor suppressor gene 5′-CpG island in breast cancer. Cancer Letter 163, 59-69.

Ohhara M, Esumi M, Kurosu Y. 1996. Activation but not inactivation of the MTS1 gene is associated with primary colorectal carcinomas. Biochemical and Biophysical Research Communication 226, 791-795.

Ovchinnikov DA, Cooper MA, Pandit P. 2011. Tumor-suppressor Gene Promoter Hypermethylation in Saliva of Head and Neck Cancer Patients. Translational oncology 5, 321-326.

Rasti M, Entezam M, Monabati A. 2009. Hypermethylation of E-Cadherin and Estrogen Receptor-a Gene Promoter and Its Association with Clinicopathological Features of Breast Cancer in Iranian Patients. International Journal of Molecular Sciences 34( 3), 50-56.

Serrano M., Hannon G J, Beach D. 1993. A new regulatory motif in cell-cycle control causing specific inhibitionof cyclin D/CDK4. Nature (Land.) 366, 704-707.

Shim YH, Kang GH, Ro JY. 2000. Correlation of p16 hypermethylation with p16 protein loss in sporadic gastric carcinomas. Laboratory Investigation 80, 689-695.

Tada T, Watanabe T, Kazama S, Kanazawa T, Hata K, Komuro Y, Nagawa H. 2003. Reduced p16 expression correlates with lymphatic invasion in colorectal cancers. Hepatogastroenterology 50, 1756-1760.

Tannapfel A, Benicke M, Katalinic A, Uhlmann D, Köckerling F, Hauss J, Wittekind C. 2001. Frequency of p16(INK4A) alterations and K-ras mutations in intrahepatic cholangiocarcinoma of the liver. Gut 47, 721-727.

Trzeciak L, Hennig E, Kolodziejski J, Nowacki M, Ostrowski J. 2001. Mutations, methylation and expression of CDKN2a/p16 gene in colorectal cancer and normal colonic mucosa. Cancer Letter 163, 17-23.

van der Riet P, Nawroz H, Hruban R H, Corio R, Tokino K, Koch w, Sidransky D. 1994. Frequent loss of chromosome 9p2l-22 early in head and neck cancer progression. Cancer Research 54, 1156-1158.

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. Clinical Cancer Res 9, 2898-2903.

Wani HA, Beigh MA, Amin S, Bhat AA, Bhat S, Khan H, Mattoo AA, Showkat M, Masood A, Majid S. 2013. Methylation profile of promoter region of p16 gene in colorectal cancer patients of Kashmir valley. Journal of Biological Regulation Homeostais Agents 27(2), 297-307.

Wiencke JK, Zheng S, Lafuente A, Lafuente MJ, Grudzen C, Wrensch MR, Miike R, Ballesta A, Trias M. 1999. Aberrant methylation of p16INK4a in anatomic and gender-specific subtypes of sporadic colorectal cancer. Cancer Epidemiological Biomarkers Prevention 8, 501-506.

Xing XB, Cai WB, Luo L, Liu LS, Shi HJ, Chen MH. 2013. The Prognostic Value of p16 Hypermethylation in Cancer: A Meta-Analysis. PLoS One 8(6), e66587.

Yi J, Wang ZW, Cang H, Chen YY, Zhao R, Yu BM, Tang XM. 2001. p16 gene methylation in colorectal cancers associated with Duke’s staging. World Journal of Gastroenterology 7, 722-725.

Yu J, Zhang HY, Sun MH, Gu J, Du X, Shi DR, Wang P, Yang ZH, Zhu JD. 2004. Methylation profile of the promoter CpG islands of 31 genes that may contribute to colorectal carcinogenesis. World Journal of Gastroenterology 10(23), 3441-54.

Zhao P, Hu YC, Talbot IC. 2003. Expressing patterns of p16 and CDK4 correlated to prognosis in colorectalcarcinoma.WorldJournalof Gastroenterology 9, 2202-2206.

Zöchbauer-Müller S, Fong KM, Virmani AK, Geradts J, Gazdar AF, Minna JD. 2001. Aberrant promoter methylation of multiple genes in non-small cell lung cancers. Cancer Research 61, 249-255.


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