Odovtos - International Journal of Dental Sciences ISSN Impreso: 1659-1046 ISSN electrónico: 2215-3411

OAI: https://revistas.ucr.ac.cr/index.php/Odontos/oai
Methylation Analysis of Rb, P16, MGMT, and hMLH-1in Mucoepidermoid Carcinomas. A Pilot Study


Methylation; Mucoepidermoid carcinoma; RB; MGMT; hMLH-1.
Metilación; Carcinoma mucoepidermoide; Rb; MGMT; hMLH-1.

How to Cite

Martínez-Marcial, L. A., Jacinto-Alemán, L. F., Trejo-Remigio, D. A., Ramírez-Martínez, C. M., & Leyva-Huerta, E. R. (2023). Methylation Analysis of Rb, P16, MGMT, and hMLH-1in Mucoepidermoid Carcinomas. A Pilot Study. Odovtos - International Journal of Dental Sciences, 25(2), 123–132. https://doi.org/10.15517/ijds.2022.53197


Mucoepidermoid carcinoma (MC) is the most common malignant epithelial neoplasm in the salivary glands. This neoplasm has varying proportions of mucous, epidermoid, intermediate, columnar, and clear cells. MCs have been associated with CRTC1-MAML2 genes; however, their pathogenesis is uncertain. Recently, epigenetic changes have been considered a possible aetiologic factor. To identify the methylation state of RB, P16, MGMT, and hMLH genes in the three severity grades of MC were used five MCs and one healthy minor salivary gland as a control group (CG) obtained from the Pathology and Oral Medicine Laboratory and analyzed using MS-PCR to compare the presence or absence of methylation in promotor regions. The Kruskal-Wallis test was performed, with p≤0.05 considered significant. CG was employed as the normalizer of methylation levels. All assays were performed in triplicate. The mean age of our population was 52.6±18.6 years old; the total population was female and included 2 low grade, 2 intermediate grade, and 1 high grade levels of severity. When comparing the methylation status of the three histopathological grades of MC against the control, statistically significant differences were observed in Rb-M, MGMT-M, and hMLH-1-NM for high-grade severity, with p values of 0.03, 0.05, and 0.04, respectively. Methylation is a possible mechanism for pathogenesis processing of high-grade MC. However, a larger sample population is necessary to validate this finding.



Granados R.H., Méndez C.M., López P.G., Aldape-Barrios B. Carcinoma mucoepidermoide. Revista de la Asociación Dental Mexicana. 2010; 67 (4): 181-4.

Avila R.E., Samar M.E., Fonseca I., Olmedo L., Asís O.G., Ferraris R. Carcinoma Mucoepidermoide de glándulas salivales: factores pronósticos histológicos e inmunohistoquímicos. International Journal of Morphology. 2011; 29 (2): 455-62.

Tronick E., Hunter R.G. Waddington, dynamic systems, and epigenetics. Front Behav Neurosci. 2016; 10: 107.

Bird A. Perceptions of epigenetics. Nature. 2007; 447 (7143): 396.

Feinberg A.P., Tycko B. The history of cancer epigenetics. Nat Rev Cancer. 2004; 4 (2): 143-53.

Jones P.A., Baylin S.B. The fundamental role of epigenetic events in cancer. Nat Rev Genet. 2002; 3 (6): 415-28.

Bird A. DNA methylation patterns and epigenetic memory. Genes Dev. 2002; 16 (1): 6-21.

Langevin S.M., Eliot M., Butler R.A., Cheong A., Zhang X., McClean M.D., et al. CpG island methylation profile in non-invasive oral rinse samples is predictive of oral and pharyngeal carcinoma. Clin Epigenetics. 2015; 7 (1): 1-12.

Rodríguez-Paredes M., Esteller M. Cancer epigenetics reaches mainstream oncology. Nat Med. 2011; 17 (3): 330-9.

Tsakogiannis D., Moschonas G.D., Daskou M., Stylianidou Z., Kyriakopoulou Z., Kottaridi C., et al. Polymorphic variability in the exon 19 of the RB1 gene and its flanking intronic sequences in HPV16-associated precancerous lesions in the Greek population. J Med Microbiol. 2018; 67 (11): 1638-44.

de Souza A.A,. Altemani A., Passador-Santos F., Turssi C.P., de Araujo N.S., de Araújo V.C., et al. Dysregulation of the Rb pathway in recurrent pleomorphic adenoma of the salivary glands. Virchows Archiv. 2015; 467 (3): 295-301.

Zhang J.C., Gao B., Yu Z.T., Liu X.B., Lu J., Xie F., et al. Promoter hypermethylation of p14 ARF, RB, and INK4 gene family in hepatocellular carcinoma with hepatitis B virus infection. Tumor Biology. 2014 Mar 1; 35 (3): 2795-802.

Guo L., Huang C., Qj J. CLINICAL STUDY Aberrant promoter hypermethylation of p16, survivin, and retinoblastoma in gastric cancer. Science Citation Index Expanded and in Journal Citation Reports/Science Edition Bratisl Med J [Internet]. 2017; 118 (3). Available from: www.elis.sk.

de Souza A.A., Altemani A., Passador-Santos F., Turssi C.P., de Araujo N.S., de Araújo V.C., et al. Dysregulation of the Rb pathway in recurrent pleomorphic adenoma of the salivary glands. Virchows Archiv. 2015; 467 (3): 295-301.

Jiao Y., Feng Y., Wang X. Regulation of tumor suppressor gene CDKN2A and encoded p16-INK4a protein by covalent modifications. Biochemistry (Moscow). 2018; 83 (11): 1289-98.

Foulkes W.D., Flanders T.Y., Pollock P.M., Hayward N.K. The CDKN2A (p16) gene and human cancer. Molecular medicine. 1997; 3 (1): 5-20.

Natsume A., Ishii D., Wakabayashi T., Tsuno T., Hatano H., Mizuno M., et al. IFN-β down-regulates the expression of DNA repair gene MGMT and sensitizes resistant glioma cells to temozolomide. Cancer Res. 2005; 65 (17): 7573-9.

Esteller M., Risques R.A., Toyota M., Capella G., Moreno V., Peinado M.A., et al. Promoter hypermethylation of the DNA repair gene O6-methylguanine-DNA methyltransferase is associated with the presence of G: C to A: T transition mutations in p53 in human colorectal tumorigenesis. Cancer Res. 2001; 61 (12): 4689-92.

Esteller M., Garcia-Foncillas J., Andion E., Goodman S.N., Hidalgo O.F., Vanaclocha V., et al. Inactivation of the DNA-repair gene MGMT and the clinical response of gliomas to alkylating agents. New England Journal of Medicine. 2000; 343 (19): 1350-4.

Guida M., Sanguedolce F., Bufo P., di Spiezio Sardo A., Bifulco G., Nappi C., et al. Aberrant DNA hypermethylation of hMLH-1 and CDKN2A/p16 genes in benign, premalignant and malignant endometrial lesions. Eur J Gynaecol Oncol [Internet]. 2009; 30 (3): 267-70. Available from: http://europepmc.org/abstract/MED/19697618

YÁÑEZ J.H.S. Análisis del Estado de metilación del ADN y su relación con Modificaciones de Histonas del promotor de los Genes MLH1, p16 y NKX2-3, en diferentes tipos de cáncer. Universidad de Concepción; 2014.

Zare M., Jafari-Nedooshan J., Jafari M., Neamatzadeh H., Abolbaghaei S.M., Foroughi E., et al. Relevance of hMLH1-93G> A, 655A> G and 1151T> A polymorphisms with colorectal cancer susceptibility: a meta-analysis based on 38 case-control studies. Rev Assoc Med Bras. 2018; 64: 942-51.

Michailidi C., Theocharis S., Tsourouflis G., Pletsa V., Kouraklis G., Patsouris E., et al. Expression and promoter methylation status of hMLH1, MGMT, APC, and CDH1 genes in patients with colon adenocarcinoma. Exp Biol Med. 2015; 240 (12): 1599-605.

Aviso de privacidad integral para pacientes. Unam.mx. [citado el 12 de agosto de 2022]. Disponible en: https://www.odonto.unam.mx/sites/default/files/inline-files/1.AP_Integral-Fac_Esc_Inst_Centros%20%20PARA%20PACIENTES%20-.pdf

Adel K. El-Naggar, Jhon K.C chan, Jennifer R. Grandis, Takashi Takata PJS. WHO Classification of Head and Neck Tumors. 4ta ed. International Aency for Research on Cancer, editor. Lyon, Francia; 2017. 347 p.

Li L.C. and Dahiya R. MethPrimer: designing primers for methylation PCRs. Bioinformatics. 2002 Nov; 18 (11): 1427-31. PMID: 12424112. MethPrimer.

Chkheidze R., Raisanen J., Gagan J., Richardson T.E., Pinho M.C., Raj K., et al. Alterations in the RB Pathway With Inactivation of RB1 Characterize Glioblastomas With a Primitive Neuronal Component. J Neuropathol Exp Neurol [Internet]. 2021 Dec 1; 80 (12): 1092-8. Available from: https://doi.org/10.1093/jnen/nlab109

Zhou Y., Jin X., Ma J., Ding D., Huang Z., Sheng H., et al. HDAC5 Loss Impairs RB Repression of Pro-Oncogenic Genes and Confers CDK4/6 Inhibitor Resistance in Cancer. Cancer Res [Internet]. 2021 Mar 15; 81 (6): 1486-99. Available from: https://doi.org/10.1158/0008-5472.CAN-20-2828

McNair C., Xu K., Mandigo A.C., Benelli M., Leiby B., Rodrigues D., et al. Differential impact of RB status on E2F1 reprogramming in human cancer. J Clin Invest [Internet]. 2018 Jan 2; 128 (1): 341-58. Available from: https://doi.org/10.1172/JCI93566

Kaelin Jr W.G. Functions of the retinoblastoma protein. Bioessays. 1999; 21 (11): 950-8.

Shieh Y., Shiah S., Jeng H., Lee H., Wu C., Chang L. DNA methyltransferase 1 expression and promoter methylation of E-cadherin in mucoepidermoid carcinoma. Cancer: Interdisciplinary International Journal of the American Cancer Society. 2005; 104 (5): 1013-21.


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