High frequency of p16 and p14 promoter hypermethylation and marked telomere instability in salivary gland tumors.
OBJECTIVE
to investigate p16(INK4a) and p14(ARF) tumor suppressor gene methylation status, determine telomere length and assess the importance of these epigenetic and genetic parameters in the development of pleomorphic adenoma and carcinoma ex pleomorphic adenoma of the parotid salivary glands.


METHODS
Genomic DNA from paraffin-embedded samples of 50 pleomorphic adenomas and 10 carcinomas ex pleomorphic adenoma was subjected to methylation specific polymerase chain reaction for hypermethylation analyses and real time polymerase chain reaction for the relative telomere length calculations.


RESULTS
Promoter hypermethylation of the two genes was a very frequent event in both neoplasms - between 60% and 90% of samples were hypermethylated - but without significant difference between the groups. The mean relative telomere length in the pleomorphic adenoma group was significantly increased in comparison to the control group (P=0.00), and significantly decreased in comparison to the carcinoma group (P=0.05). Telomeres were also longer in myxoid and cellular histological subtypes of adenomas than in the classic type (P=0.044 and P=0.018, respectively). Longer telomeres were more frequent in tumors with hypermethylated p14(ARF) alleles (P=0.013).


CONCLUSIONS
Promoter hypermethylations seems to be an important mechanism of p16(INK4a) and p14(ARF) inactivation in parotid gland tumors. Telomeric lengthening appears to be involved in the pathogenesis of both benign and malignant tumors of the parotid glands.
Institute of Human Genetics, School of Dental Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia.
Nadja
N
Nikolic
Clinic for Maxillofacial Surgery, School of Dental Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia.
Boban
B
Anicic
Institute of Human Genetics, School of Dental Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia.
Jelena
J
Carkic
Institute of Human Genetics, School of Dental Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia.
Jelena
J
Simonovic
Institute of Human Genetics, School of Dental Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia.
Bosko
B
Toljic
Laboratory for Radiobiology and Molecular Genetics, Institute of Nuclear Sciences "Vinca", University of Belgrade, PO box 522, 11000 Belgrade, Serbia.
Nasta
N
Tanic
Institute of Pathology, School of Dental Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia.
Zvezdana
Z
Tepavcevic
Clinic for Maxillofacial Surgery, School of Dental Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia.
Miroslav
M
Vukadinovic
Clinic for Maxillofacial Surgery, School of Dental Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia.
Vitomir S
VS
Konstantinovic
Institute of Human Genetics, School of Dental Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia. Electronic address: jelena.milasin@stomf.bg.ac.rs.
Jelena
J
Milasin
D008949:Adenoma, Pleomorphic / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D000328:Adult; D000368:Aged; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D005260:Female; D016147:Genes, Tumor Suppressor; D019942:Genes, p16; D006801:Humans; D008297:Male; D008875:Middle Aged; D060787:Neoplasm Grading; D015513:Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D010307:Parotid Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D011401:Promoter Regions, Genetic; D016615:Telomere / Q000235:genetics* / Q000378:metabolism; D059505:Telomere Homeostasis; D055815:Young Adult
C401561:CDK2AP2 protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D015513:Oncogene Proteins
Parotid gland tumors; Promoter hypermethylation; Telomere length; p14; p16