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Establishment and characterization of a primary calcifying epithelial odontogenic tumor cell population
Author(s) -
Amm Hope M.,
Rollins Douglas L.,
Ren Changchun,
Dong Juan,
DeVilliers Patricia,
Rivera Helen,
MacDougall Mary
Publication year - 2014
Publication title -
journal of oral pathology and medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.887
H-Index - 83
eISSN - 1600-0714
pISSN - 0904-2512
DOI - 10.1111/jop.12125
Subject(s) - ptch1 , biology , pathology , odontogenic tumor , population , adamantinoma , cancer research , microbiology and biotechnology , ameloblastoma , gene , genetics , medicine , maxilla , anatomy , odontogenic , sonic hedgehog , environmental health
Calcifying epithelial odontogenic tumors ( CEOT s) are rare neoplasms derived from dental tissue with the unique characteristic of calcifying amyloid‐like material. Objectives To establish primary CEOT epithelial‐derived cell populations, investigate the expression of enamel matrix proteins ( EMP s), and identify potential ameloblastin ( AMBN ) and patched 1 ( PTCH 1 ) gene alterations. Materials and Methods A 28‐year‐old patient with a lesion of the posterior maxilla, radiographically characterized by a radiolucency with well‐defined borders containing mixed radiopacities, agreed to participate with informed consent. The patient's biopsy confirmed the diagnosis of CEOT , and a small representative tumor fragment was ascertained for cell culture. Explant cultures were established and used to establish primary cell populations. These were analyzed for morphology, cell proliferation, mineralization activity, expression of epithelial‐associated markers ( qRT ‐PCR and immunocytochemistry), and gene mutations of AMBN or PTCH1 . DNA was extracted from tumor cells and gene coding and exon–intron boundaries overlapping fragments amplified. PCR products were bidirectional DNA sequenced and compared against reference sequence. Results A CEOT cell population was established and proliferated in culture and could be maintained for several passages. Expression of EMP s, cytokeratin 14 and 17, and patched ( PTCH1 ), as well as ALP activity, was detected. These cells also had the ability to mineralize, similar to the primary tumor. Two AMBN alterations were identified in the sample: c.1323G>A/A441A (rs7680880) and c.1344*+111delA. Two single‐nucleotide polymorphisms were identified in the PTCH1 gene. Conclusions Our data support the establishment of a CEOT ‐derived cell population, which expresses known epithelial‐associated proteins.