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9-methoxycamptothecin from Nothapodytes foetida induces apoptosis in murine sarcoma S180 cells
Na Liao1, Peng Zhang, Mingzhang Ao
1Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
Abstract:
9-Methoxycamptothecin (MCPT) was found to have antitumour activities through topoisomerase inhibition. However, the type of cell death induced in the tumour cells treated with MCPT was not elucidated. In this study, MCPT and camptothecin were isolated from Nothapodytes foetida distributed in Hubei Province, China and identified by NMR spectroscopy. MCPT was tested by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay using camptothecin as reference. Annexin V-FITC/propidium iodide double staining and real-time PCR were also performed. The IC50 value was (0.385 +/- 0.08) microM. The apoptosis rates increased from 9.5% to 17.27%, 30.14%, and 66.46% with an increase in MCPT concentrations from 0, 0.19, 0.38, to 0.95 microM, respectively. The ratio of Bax/Bcl-2 also increased from 1 to 1.61, 2.43, and 4.57, respectively. Bax and Bcl-2 are crucial to the mitochondria pathway. The results indicate that the mitochondria pathway may be involed in MCPT-induced murine sarcoma S180 apoptosis.
Insights
9-Methoxycamptothecin (MCPT) induces apoptosis in murine sarcoma S180 cells. This study reveals that the mitochondria pathway is involved in MCPT
Area of Science:
- Pharmacology
- Molecular Biology
- Cancer Research
Background:
- 9-Methoxycamptothecin (MCPT) exhibits antitumour properties via topoisomerase inhibition.
- The specific mechanism of cell death induced by MCPT in tumour cells remained unclear.
- Understanding MCPT's cellular effects is crucial for its therapeutic development.
Purpose of the Study:
- To investigate the type of cell death induced by 9-Methoxycamptothecin (MCPT) in tumour cells.
- To elucidate the molecular pathways involved in MCPT-mediated apoptosis.
- To determine the efficacy of MCPT compared to camptothecin.
Main Methods:
- Isolation and identification of MCPT and camptothecin from Nothapodytes foetida using NMR spectroscopy.
- Antitumour activity assessment using the MTT assay and Annexin V-FITC/propidium iodide double staining.
- Analysis of apoptosis-related gene expression (Bax and Bcl-2) via real-time PCR.
Main Results:
- MCPT demonstrated significant antitumour activity with an IC50 value of (0.385 +/- 0.08) microM.
- MCPT treatment dose-dependently increased apoptosis rates in S180 cells.
- The Bax/Bcl-2 ratio increased significantly, indicating activation of the mitochondrial apoptosis pathway.
Conclusions:
- 9-Methoxycamptothecin (MCPT) induces apoptosis in murine sarcoma S180 cells.
- The mitochondrial pathway plays a key role in MCPT-induced apoptosis.
- MCPT holds potential as an anticancer agent, warranting further investigation.
