Related Experiment Video
Updated: Oct 11, 2026
Studying Cell Cycle-regulated Gene Expression by Two Complementary Cell Synchronization Protocols
Published on: June 6, 2017
High pressure sensitizes murine erythroleukemia cells to caffeine-induced premature mitosis
M Matsumoto1, T Yamaguchi, K Nakazono
1Department of Chemistry, Faculty of Science, Fukuoka University, Fukuoka, 814-0180, Japan. takeo@fukuoka-u.ac.jp
Abstract:
Murine erythroleukemia (MEL) cells were exposed to a high pressure of 80 MPa or aphidicolin (APH), DNA polymerase inhibitor. The effects of caffeine on cell cycle were examined using these cells. During the culture of 80 MPa-treated MEL cells at atmospheric pressure, the cells arrested in the G2 phase, and cyclin B and hyperphosphorylated p34(cdc2) were accumulated. Namely, maturation promoting factor (MPF) composed of p34(cdc2) and cyclin B was inactive. However, upon exposure to caffeine, these cells entered prematurely into mitosis by activating MPF. Caffeine-induced premature mitosis was suppressed by butyrolactone I and orthovanadate. On the other hand, APH-treated MEL cells, which were not exposed to 80 MPa, were not so sensitive to caffeine-induced premature mitosis despite cyclin B accumulation. In this case, dephosphorylation of p34(cdc2) was not induced by caffeine. Interestingly, caffeine-induced premature mitosis in the 80 MPa-treated cells was also suppressed by APH. These results suggest that the premature mitosis of 80 MPa-treated MEL cells by caffeine is induced by active MPF, and that APH-sensitive molecules such as DNA polymerase may also play an important role in the checkpoint that controls the transition from G2 to M phase.
Insights
Caffeine triggers premature mitosis in high-pressure-treated cells by activating maturation promoting factor (MPF). Aphidicolin (APH), a DNA polymerase inhibitor, also suppresses this caffeine-induced effect, highlighting its role in cell cycle checkpoints.
Area of Science:
- Cell Biology
- Molecular Biology
- Biophysics
Background:
- High pressure (80 MPa) and aphidicolin (APH) induce cell cycle arrest in murine erythroleukemia (MEL) cells.
- Cells arrest in G2 phase with accumulated cyclin B and hyperphosphorylated p34(cdc2), indicating inactive maturation promoting factor (MPF).
Purpose of the Study:
- To investigate the effects of caffeine on cell cycle progression in MEL cells exposed to high pressure or APH.
- To elucidate the role of MPF activation and DNA polymerase in the G2/M checkpoint control.
Main Methods:
- Murine erythroleukemia (MEL) cells were treated with 80 MPa high pressure or aphidicolin (APH).
- Caffeine was administered to assess its impact on cell cycle progression and MPF activity.
- Inhibitors like butyrolactone I and orthovanadate were used to study suppression mechanisms.
Main Results:
- Caffeine induced premature mitosis in 80 MPa-treated cells by activating MPF, an effect suppressed by butyrolactone I and orthovanadate.
- APH-treated cells showed less sensitivity to caffeine-induced premature mitosis, with no p34(cdc2) dephosphorylation.
- Aphidicolin (APH) suppressed caffeine-induced premature mitosis in 80 MPa-treated cells, suggesting a role for DNA polymerase in the G2/M checkpoint.
Conclusions:
- Caffeine promotes premature mitosis in high-pressure-treated MEL cells via MPF activation.
- APH-sensitive molecules, potentially DNA polymerase, are crucial for the G2/M phase transition checkpoint.
More Related Videos
09:09Preparation of Primary Acute Lymphoblastic Leukemia Cells in Different Cell Cycle Phases by Centrifugal Elutriation
Published on: November 10, 2017
06:00Through the Looking Glass: Time-lapse Microscopy and Longitudinal Tracking of Single Cells to Study Anti-cancer Therapeutics
Published on: May 14, 2016
