TCF21hypermethylation regulates renal tumor cell clonogenic proliferation and migration

Autor: Saskia L. Gooskens, Mary M. van den Heuvel-Eibrink, Robert Pieters, Timothy D. Klasson, Elizabeth J. Perlman, Rachel H. Giles, Hendrik Gremmels, Ive Logister
Přispěvatelé: Pediatrics
Rok vydání: 2017
Předmět:
0301 basic medicine
renal cell carcinoma
Cancer Research
Epithelial-Mesenchymal Transition
tumor suppressor
Vimentin
epithelial‐to‐mesenchymal transition
lcsh:RC254-282
03 medical and health sciences
Antigens
CD
Cell Movement
Cell Line
Tumor
Basic Helix-Loop-Helix Transcription Factors
Journal Article
Genetics
medicine
Humans
Epithelial–mesenchymal transition
Clonogenic assay
Carcinoma
Renal Cell
Research Articles
Tumor Stem Cell Assay
Cell Proliferation
TCF21
biology
Cell growth
General Medicine
DNA Methylation
Cadherins
clear cell sarcoma of the kidney
lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens
medicine.disease
Molecular biology
Kidney Neoplasms
Clear cell renal cell carcinoma
030104 developmental biology
Oncology
Cancer cell
SNAI1
biology.protein
Cancer research
Molecular Medicine
Ectopic expression
Snail Family Transcription Factors
epithelial-to-mesenchymal transition
methylation
Research Article
Zdroj: Molecular Oncology, 12(2), 166-179. John Wiley & Sons Ltd.
Molecular Oncology, Vol 12, Iss 2, Pp 166-179 (2018)
Molecular Oncology, 12(2), 166. Elsevier
Molecular Oncology
ISSN: 1574-7891
DOI: 10.1002/1878-0261.12149
Popis: We recently identified hypermethylation at the gene promoter of transcription factor 21 (TCF21) in clear cell sarcoma of the kidney (CCSK), a rare pediatric renal tumor. TCF21 is a transcription factor involved in tubular epithelial development of the kidney and is a candidate tumor suppressor. As there are no in vitro models of CCSK, we employed a well-established clear cell renal cell carcinoma (ccRCC) cell line, 786-O, which also manifests high methylation at the TCF21 promoter, with consequent low TCF21 expression. The tumor suppressor function of TCF21 has not been functionally addressed in ccRCC cells; we aimed to explore the functional potential of TCF21 expression in ccRCC cells in vitro. 786-O clones stably transfected with either pBABE-TCF21-HA construct or pBABE vector alone were functionally analyzed. We found that ectopic expression of TCF21 in 786-O cells results in a trend toward decreased cell proliferation (not significant) and significantly decreased migration compared with mock-transfected 786-O cells. Although the number of colonies established in colony formation assays was not different between 786-O clones, colony size was significantly reduced in 786-O cells expressing TCF21. To investigate whether the changes in migration were due to epithelial-to-mesenchymal transition changes, we interrogated the expression of selected epithelial and mesenchymal markers. Although we observed upregulation of mRNA and protein levels of epithelial marker E-cadherin in clones overexpressing TCF21, this did not result in surface expression of E-cadherin as measured by fluorescence-activated cell sorting and immunofluorescence. Furthermore, mRNA expression of the mesenchymal markers vimentin (VIM) and SNAI1 was not significantly decreased in TCF21-expressing 786-O cells, while protein levels of VIM were markedly decreased. We conclude that re-expression of TCF21 in renal cancer cells that have silenced their endogenous TCF21 locus through hypermethylation results in reduced clonogenic proliferation, reduced migration, and reduced mesenchymal-like characteristics, suggesting a tumor suppressor function for transcription factor 21.
Databáze: OpenAIRE
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