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Glucosylceramide synthase and its functional interaction with RTN-1C regulate chemotherapeutic-induced apoptosis in
Federica Di Sano1, Barbara Fazi, Gennaro Citro
1Department of Biology, University of Rome Tor Vergata, 00133 Rome, Italy.
Abstract:
Glucosylceramide synthase (GCS), the key enzyme in the biosynthesis of glycosphingolipids, has been implicated in many biological phenomena, including multidrug resistance. GCS inhibition, by both antisense and the specific inhibitor (D-threo)-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), results in a drastic decrease of apoptosis induced by the p53-independent chemotherapeutic agent N-(4-hydroxyphenyl)retinamide in neuroepithelioma cells. By using the yeast two-hybrid system, we have identified a member of the reticulon (RTN) family (RTN-1C) as the major GCS-protein partner. Interestingly, RTN-1C not only interacts with GCS at Golgi/ER interface but also modulates its catalytic activity in situ. In fact, overexpression of RTN-1C sensitizes CHP-100 cells to fenretinide-induced apoptosis. These findings demonstrate a novel p53-independent pathway of apoptosis regulated by Golgi/endoplasmic reticulum protein interactions, which is relevant for cancer combined therapy.
Insights
Glucosylceramide synthase (GCS) inhibition reduces chemotherapy-induced apoptosis. A novel protein interaction between GCS and reticulon-1C (RTN-1C) regulates this pathway, offering new avenues for cancer therapy.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Glucosylceramide synthase (GCS) is crucial for glycosphingolipid synthesis and implicated in multidrug resistance.
- GCS inhibition affects apoptosis induced by certain chemotherapeutic agents.
- Understanding GCS regulation is vital for developing effective cancer treatments.
Purpose of the Study:
- To identify GCS-interacting proteins.
- To elucidate the role of GCS-protein interactions in apoptosis.
- To explore novel therapeutic strategies for cancer.
Main Methods:
- Yeast two-hybrid system for protein interaction screening.
- Use of GCS inhibitors like PDMP and antisense strategies.
- Overexpression of identified interacting proteins in cancer cell lines.
- Assessment of apoptosis induction by chemotherapeutic agents.
Main Results:
- Identified reticulon-1C (RTN-1C) as a major GCS-binding protein.
- RTN-1C interacts with GCS at the Golgi/ER interface.
- RTN-1C modulates GCS activity in situ.
- Overexpression of RTN-1C sensitizes cells to fenretinide-induced apoptosis.
Conclusions:
- Discovered a novel p53-independent apoptosis pathway involving Golgi/ER protein interactions.
- RTN-1C is a key regulator of GCS function in apoptosis.
- This pathway represents a potential target for combined cancer therapy.
