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Updated: Jun 25, 2025
Prediction and Validation of Gene Regulatory Elements Activated During Retinoic Acid Induced Embryonic Stem Cell Differentiation
Published on: June 21, 2016
Stemness and Cell Cycle Regulators and Their Modulation by Retinoic Acid in Ewing Sarcoma
Maria Eduarda Battistella1, Natália Hogetop Freire1,2,3, Bruno Toson1
1Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, Porto Alegre 90035-003, Brazil.
Abstract:
Retinoic acid (RA) regulates stemness and differentiation in human embryonic stem cells (ESCs). Ewing sarcoma (ES) is a pediatric tumor that may arise from the abnormal development of ESCs. Here we show that RA impairs the viability of SK-ES-1 ES cells and affects the cell cycle. Cells treated with RA showed increased levels of p21 and its encoding gene, CDKN1A. RA reduced mRNA and protein levels of SRY-box transcription factor 2 (SOX2) as well as mRNA levels of beta III Tubulin (TUBB3), whereas the levels of CD99 increased. Exposure to RA reduced the capability of SK-ES-1 to form tumorspheres with high expression of SOX2 and Nestin. Gene expression of CD99 and CDKN1A was reduced in ES tumors compared to non-tumoral tissue, whereas transcript levels of SOX2 were significantly higher in tumors. For NES and TUBB3, differences between tumors and control tissue did not reach statistical significance. Low expression of CD99 and NES, and high expression of SOX2, were significantly associated with a poorer patient prognosis indicated by shorter overall survival (OS). Our results indicate that RA may display rather complex modulatory effects on multiple target genes associated with the maintenance of stem cell's features versus their differentiation, cell cycle regulation, and patient prognosis in ES.
Insights
Retinoic acid (RA) impacts Ewing sarcoma (ES) cell viability and stemness markers like SOX2. RA
Area of Science:
- Developmental Biology
- Cancer Research
- Stem Cell Biology
Background:
- Retinoic acid (RA) is crucial for regulating stemness and differentiation in human embryonic stem cells (ESCs).
- Ewing sarcoma (ES), a pediatric tumor, may originate from aberrant ESC development.
- Understanding RA's role in ES could reveal therapeutic targets.
Purpose of the Study:
- To investigate the effects of retinoic acid (RA) on Ewing sarcoma (ES) cell line SK-ES-1.
- To analyze RA's impact on stemness markers, cell cycle, and tumor formation.
- To correlate gene expression patterns with patient prognosis in ES.
Main Methods:
- Treatment of SK-ES-1 cells with RA.
- Analysis of cell viability, cell cycle, and tumorsphere formation.
- Quantitative PCR and Western blotting for gene and protein expression (SOX2, TUBB3, CD99, CDKN1A, NES).
- Correlation of gene expression with clinical data and overall survival (OS).
Main Results:
- RA impaired SK-ES-1 cell viability and altered cell cycle, increasing p21 (CDKN1A) levels.
- RA reduced SOX2 and TUBB3 expression but increased CD99 expression.
- RA diminished tumorsphere formation capacity and SOX2/Nestin expression.
- In ES tumors, SOX2 was upregulated, while CD99 and CDKN1A were downregulated compared to normal tissue.
- Low CD99/NES and high SOX2 expression correlated with poorer patient prognosis (shorter OS).
Conclusions:
- RA exhibits complex modulatory effects on genes involved in stemness, differentiation, and cell cycle regulation in ES.
- Specific gene expression patterns (SOX2, CD99, NES) influenced by RA are linked to ES prognosis.
- RA's intricate role warrants further investigation for potential therapeutic strategies in ES.
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