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The molecular mechanisms of 5-fluoro-2'-deoxyuridine induced cell death
A Nagano1, T Kakutani, Y Matumoto
1Faculty of Pharmaceutical Sciences, Okayama University, Japan.
Abstract:
The molecular mechanism of cell death induced by 5-Fluoro-2'-deoxyuridine (FUdR) was investigated. FUdR caused cell death to induce dNTP pool imbalance and following DNA double strand breaks in mouse mammary tumor FM3A cells. We isolated a new endonuclease from FUdR-treated cells, named endonuclease S, that played an important role in FUdR-induced cell death. Cells treated with FUdR showed intracellular acidification before cell death formation. We observed that the endonuclease S in acidic cells may lead the DNA fragmentation. On the other hand, we observed that protease inhibitors (such as TLCK, TPCK, PMSF, p-APMSF, Pefabloc SC and Z-Asp-CH2-DCB) blocked intracellular acidification, DNA fragmentation and FUdR-induced cell death. But the inhibitors did not affect dNTP pool imbalance in the cells. These results suggest that proteases act at the point of downstream of dNTP pool imbalance and upstream of the intracellular acidification.
Insights
5-Fluoro-2'-deoxyuridine (FUdR) triggers cell death by disrupting DNA and activating endonuclease S. Protease inhibitors block this process, suggesting proteases are key downstream mediators in FUdR-induced apoptosis.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- 5-Fluoro-2 -deoxyuridine (FUdR) is a chemotherapeutic agent.
- The precise molecular mechanisms underlying FUdR-induced cell death require further elucidation.
Purpose of the Study:
- To investigate the molecular mechanism of cell death induced by 5-Fluoro-2 -deoxyuridine (FUdR).
- To identify key enzymes and cellular events involved in FUdR-induced apoptosis.
Main Methods:
- Investigated FUdR effects on dNTP pools and DNA integrity in mouse mammary tumor FM3A cells.
- Isolated and characterized a novel endonuclease (endonuclease S) from FUdR-treated cells.
- Utilized protease inhibitors to probe the role of proteases in FUdR-induced cell death.
Main Results:
- FUdR induced dNTP pool imbalance and DNA double-strand breaks, leading to cell death.
- A novel endonuclease S was identified, playing a crucial role in FUdR-induced DNA fragmentation.
- Intracellular acidification preceded cell death, and endonuclease S activity in acidic conditions was implicated in DNA fragmentation.
- Protease inhibitors blocked intracellular acidification, DNA fragmentation, and cell death, without affecting dNTP pool imbalance.
Conclusions:
- Proteases act downstream of dNTP pool imbalance and upstream of intracellular acidification in the FUdR-induced cell death pathway.
- Endonuclease S activation in an acidic cellular environment is a critical step in FUdR-induced DNA fragmentation and apoptosis.
- Protease activity is essential for the progression of FUdR-induced cell death, highlighting potential therapeutic targets.