PUBLICATION
TCF21 hypermethylation regulates renal tumor cell clonogenic proliferation and migration
- Authors
- Gooskens, S.L., Klasson, T.D., Gremmels, H., Logister, I., Pieters, R., Perlman, E.J., Giles, R.H., van den Heuvel-Eibrink, M.M.
- ID
- ZDB-PUB-190207-11
- Date
- 2018
- Source
- Molecular Oncology 12: 166-179 (Journal)
- Registered Authors
- Logister, Ive
- Keywords
- TCF21, clear cell sarcoma of the kidney, epithelial-to-mesenchymal transition, methylation, renal cell carcinoma, tumor suppressor
- MeSH Terms
-
- Antigens, CD/genetics
- Antigens, CD/metabolism
- Basic Helix-Loop-Helix Transcription Factors/genetics
- Basic Helix-Loop-Helix Transcription Factors/metabolism*
- Cadherins/genetics
- Cadherins/metabolism
- Carcinoma, Renal Cell/genetics*
- Carcinoma, Renal Cell/metabolism
- Cell Line, Tumor
- Cell Movement/genetics*
- Cell Proliferation/genetics*
- DNA Methylation/genetics*
- Epithelial-Mesenchymal Transition
- Humans
- Kidney Neoplasms/genetics*
- Kidney Neoplasms/metabolism
- Snail Family Transcription Factors/genetics
- Snail Family Transcription Factors/metabolism
- Tumor Stem Cell Assay
- Vimentin/genetics
- Vimentin/metabolism
- PubMed
- 29080283 Full text @ Mol. Oncol.
Citation
Gooskens, S.L., Klasson, T.D., Gremmels, H., Logister, I., Pieters, R., Perlman, E.J., Giles, R.H., van den Heuvel-Eibrink, M.M. (2018) TCF21 hypermethylation regulates renal tumor cell clonogenic proliferation and migration. Molecular Oncology. 12:166-179.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping