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2015 ; 23
(4
): 663-80
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A novel canine kidney cell line model for the evaluation of neoplastic
development: karyotype evolution associated with spontaneous immortalization and
tumorigenicity
#MMPMID25957863
Omeir R
; Thomas R
; Teferedegne B
; Williams C
; Foseh G
; Macauley J
; Brinster L
; Beren J
; Peden K
; Breen M
; Lewis AM Jr
Chromosome Res
2015[Dec]; 23
(4
): 663-80
PMID25957863
show ga
The molecular mechanisms underlying spontaneous neoplastic transformation in
cultured mammalian cells remain poorly understood, confounding recognition of
parallels with the biology of naturally occurring cancer. The broad use of
tumorigenic canine cell lines as research tools, coupled with the accumulation of
cytogenomic data from naturally occurring canine cancers, makes the domestic dog
an ideal system in which to investigate these relationships. We developed a
canine kidney cell line, CKB1-3T7, which allows prospective examination of the
onset of spontaneous immortalization and tumorigenicity. We documented the
accumulation of cytogenomic aberrations in CKB1-3T7 over 24 months in continuous
culture. The majority of aberrations emerged in parallel with key phenotypic
changes in cell morphology, growth kinetics, and tumor incidence and latency.
Focal deletion of CDKN2A/B emerged first, preceding the onset and progression of
tumorigenic potential, and progressed to a homozygous deletion across the cell
population during extended culture. Interestingly, CKB1-3T7 demonstrated a
tumorigenic phenotype in vivo prior to exhibiting loss of contact inhibition in
vitro. We also performed the first genome-wide characterization of the canine
tumorigenic cell line MDCK, which also exhibited CDKN2A/B deletion. MDCK and
CKB1-3T7 cells shared several additional aberrations that we have reported
previously as being highly recurrent in spontaneous canine cancers, many of
which, as with CDKN2A/B deletion, are evolutionarily conserved in their human
counterparts. The conservation of these molecular events across multiple species,
in vitro and in vivo, despite their contrasting karyotypic architecture, is a
powerful indicator of a common mechanism underlying emerging neoplastic activity.
Through integrated cytogenomic and phenotypic characterization of serial passages
of CKB1-3T7 from initiation to development of a tumorigenic phenotype, we present
a robust and readily accessible model (to be made available through the American
Type Culture Collection) of spontaneous neoplastic transformation that overcomes
many of the limitations of earlier studies.