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Updated: Jun 26, 2026
Establishing Cell Lines Overexpressing DR3 to Assess the Apoptotic Response to Anti-mitotic Therapeutics
Published on: January 11, 2019
Reactivation of death receptor 4 (DR4) expression sensitizes medulloblastoma cell lines to TRAIL
Dolly G Aguilera1, Chandra M Das, Neeta D Sinnappah-Kang
1Department of Pediatrics, University of Texas, M.D. Anderson Cancer Center, Houston, 77030, USA.
Object:
Apoptosis, a key cellular response to therapeutic agents is often inactivated in tumor cells. In this study, we evaluated the expression of the tumor necrosis family of death receptors, DR4 and DR5, in medulloblastoma tumor samples and cell lines to determine if epigenetic modulation of gene expression could sensitize tumor cell lines to TRAIL-mediated apoptosis.
Methods:
Human medulloblastoma samples and cell lines were analyzed for DR4 and DR5 expression by quantitative PCR and immunofluorescence assays. Cell lines with downregulated expression of one or both genes were treated with the histone deacetylase inhibitor, MS-275, and the expression of DR4 and DR5 measured by quantitative PCR, Western blotting, flow cytometry and chromatin immunoprecipitation assays. Induction of apoptosis in the presence of MS-275 was evaluated by TUNEL assay and its ability to augment TRAIL-mediated cytotoxicity was determined by MTT assays, Western blotting and flow cytometry.
Results:
Compared to normal cerebellum, DR4, but not DR5 expression was consistently downregulated in medulloblastoma tumor samples and in Daoy and D283 cell lines. Interestingly, MS-275 decreased cell growth and induced apoptosis in Daoy and D283 cells. In Daoy cells, this coincided with increased histone H3 and H4 acetylation at the DR4 promoter and enhanced DR4 gene and protein expression as well as elevated Caspase-8 activity. The involvement of DR4 in the cellular response to MS-275 was further confirmed by the observation that knockdown of DR4 and FADD abrogated apoptosis. Further, addition of TRAIL to MS-275 treated cells resulted in an enhancement of apoptosis, suggesting that the upregulated death receptors were functional.
Conclusion:
Our study provides an understanding of the role of DR4 in apoptosis of medulloblastoma cell lines and suggests a potential contribution of aberrant histone deacetylation to the resistance of medulloblastoma cells to therapeutic death.
Insights
Epigenetic drug MS-275 upregulates DR4 (death receptor 4) in medulloblastoma cells, enhancing TRAIL-mediated apoptosis and overcoming resistance. This suggests targeting histone deacetylation could be a therapeutic strategy.
Area of Science:
- Oncology
- Molecular Biology
- Epigenetics
Background:
- Apoptosis is crucial for cancer therapy but often impaired in medulloblastoma.
- Tumor necrosis family death receptors, DR4 and DR5, mediate apoptosis.
- Epigenetic silencing can downregulate death receptor expression in tumors.
Purpose of the Study:
- To investigate DR4 and DR5 expression in medulloblastoma.
- To determine if epigenetic modulation can restore TRAIL-mediated apoptosis.
- To explore the role of histone deacetylase inhibition in medulloblastoma cell sensitization.
Main Methods:
- Quantitative PCR and immunofluorescence for DR4/DR5 expression analysis.
- Treatment with histone deacetylase inhibitor MS-275.
- Assays for apoptosis induction, gene/protein expression, and histone acetylation.
Main Results:
- DR4 was downregulated in medulloblastoma samples and cell lines.
- MS-275 treatment decreased cell growth and induced apoptosis.
- MS-275 increased DR4 expression and sensitized cells to TRAIL-induced apoptosis.
Conclusions:
- Aberrant histone deacetylation contributes to medulloblastoma's resistance to apoptosis.
- DR4 plays a significant role in MS-275-induced apoptosis in medulloblastoma.
- Targeting epigenetic modifications offers a potential therapeutic avenue for medulloblastoma.
