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Pleiotropic effects of fenretinide in neuroblastoma cell lines and multicellular tumor spheroids
Roos Cuperus1, Godelieve A M Tytgat, René Leen
1Laboratory Genetic Metabolic Diseases and Department of Pediatrics/Emma Children's Hospital, Academic Medical Centre, University of Amsterdam, 1100 DE Amsterdam, The Netherlands.
Abstract:
The efficacy and mechanism of action of fenretinide (4-HPR), a vitamin A analogue, was investigated in a panel of six neuroblastoma cell lines and multicellular tumor spheroids. The latter are three dimensional cell aggregates and as such, a model for micrometastases. In all cell lines, the production of reactive oxygen species (ROS) increased with 163-680% after 1 h of treatment with 4-HPR. In addition, a decrease of the mitochondrial membrane potential of 30-75% was observed after 4 h of incubation with 4-HPR. A 6-12-fold difference was observed between the IC50 values for cell proliferation and viability between the most sensitive (IMR32) and most resistant (NASS) cell line towards 4-HPR. Flow cytometric analysis showed an increased amount of apoptotic bodies and no cell-cycle arrest. The antioxidant Trolox completely inhibited the accumulation of 4HPR-induced ROS and prevented the 4HPR-associated cytotoxicity. In all neuroblastoma spheroids, 4-HPR induced a complete cytostasis at clinical relevant concentrations (3-10 microM). Immunohistochemical analysis of 4-HPR-treated spheroids showed a decreased staining for proliferation marker Ki-67 and an increased staining for cleaved-PARP, a marker of apoptosis. Our results suggest that 4-HPR might be a promising agent for the treatment of micrometastases and high-risk neuroblastoma.
Insights
Fenretinide (4-HPR), a vitamin A analogue, effectively combats neuroblastoma by increasing reactive oxygen species and inducing apoptosis. This agent shows promise for treating high-risk neuroblastoma and micrometastases.
Area of Science:
- Oncology
- Pharmacology
- Cell Biology
Background:
- Neuroblastoma is a pediatric cancer with high-risk subtypes and micrometastases.
- Fenretinide (4-HPR), a vitamin A derivative, is being investigated for its anti-cancer properties.
Purpose of the Study:
- To evaluate the efficacy and mechanism of action of fenretinide (4-HPR) in neuroblastoma.
- To assess 4-HPR's effect on neuroblastoma cell lines and multicellular tumor spheroids, a model for micrometastases.
Main Methods:
- Treatment of six neuroblastoma cell lines and multicellular tumor spheroids with 4-HPR.
- Measurement of reactive oxygen species (ROS) production and mitochondrial membrane potential.
- Analysis of cell proliferation, viability, apoptosis, and cell-cycle progression using flow cytometry and immunohistochemistry.
Main Results:
- 4-HPR significantly increased ROS production (163-680%) and decreased mitochondrial membrane potential (30-75%) in neuroblastoma cells.
- Significant differences in IC50 values were observed between sensitive (IMR32) and resistant (NASS) cell lines.
- 4-HPR induced apoptosis, evidenced by increased apoptotic bodies and cleaved-PARP, without causing cell-cycle arrest.
- 4-HPR demonstrated complete cytostasis in neuroblastoma spheroids at clinically relevant concentrations (3-10 microM).
Conclusions:
- Fenretinide (4-HPR) exhibits potent anti-neuroblastoma activity through ROS generation and apoptosis induction.
- The antioxidant Trolox mitigated 4-HPR's cytotoxic effects, confirming the role of ROS.
- 4-HPR is a promising therapeutic agent for high-risk neuroblastoma and micrometastases.
