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In vitro mitochondrial apoptosis of melanoma cells via immature Poncirus trifoliata fruit extract
1Research Institute of Clinical Medicine of Jeonbuk National University - Biomedical Research Institute of Jeonbuk National University Hospital, Jeonju, Korea. children@jbnu.ac.kr.
Objective:
Poncirus trifoliata (P. trifoliata) fruits exert phytotherapeutic effects, depending on their maturity level. However, the mechanism by which these phytotherapeutic effects are exerted remains undefined - especially in cancers. Therefore, in this study, we investigated the effects of the immature fruit extract of P. trifoliata on a B16 melanoma cell line.
Materials And Methods:
The effect of immature P. trifoliata extract on B16 cells was evaluated by MTT assay, cell proliferation, FACScan analysis of cell cycles, confocal imaging analysis, nuclear (Hoechst) staining, apoptosis assay (Annexin V-fluorescein isothiocyanate/propidium iodide staining), and Western blot assay. The capacity of immature P. trifoliata extract to inhibit the invasion and migration of B16 cells was assessed using the scratch-wound assay and Matrigel migration assay. The effect of immature P. trifoliata extract on mitochondrial function was determined via the mitochondrial membrane potential assay, activity, and fraction and cytosol proteins.
Results:
Treating B16 cells with a methanol extract of immature P. trifoliata (MEPT) significantly inhibited cell viability, migration, and invasiveness in a dose- (p<0.01) and time (p<0.01)- dependent manner. MEPT arrested the cells in the G1 phase of the cell cycle and led to the activation of the PI3K/AKT/p21 pathway. Furthermore, MEPT dose-dependently induced apoptosis in B16 cells by increasing the expression of the pro-apoptotic proteins Bax and Apaf-1, while decreasing the expression of the anti-apoptotic protein, Bcl-2. MEPT treatment also decreased mitochondrial membrane potential.
Conclusions:
Immature P. trifoliata extract inhibited the growth of melanoma cells by inducing cell apoptosis through mitochondrial pathways. Therefore, further research into immature P. trifoliata extract as a potential therapeutic compound for melanoma treatment is warranted.
Insights
Immature Poncirus trifoliata fruit extract effectively inhibits melanoma cell growth by inducing apoptosis via mitochondrial pathways. This study highlights its potential as a natural therapeutic for melanoma.
Area of Science:
- Phytotherapy
- Cancer Research
- Molecular Biology
Background:
- Poncirus trifoliata (P. trifoliata) fruits possess phytotherapeutic properties influenced by maturity.
- The precise mechanisms underlying P. trifoliata's effects, particularly in cancer, are not fully understood.
- This study focuses on the anti-cancer potential of immature P. trifoliata fruit extract against melanoma.
Purpose of the Study:
- To investigate the effects of immature P. trifoliata fruit extract on B16 melanoma cells.
- To elucidate the molecular mechanisms by which the extract inhibits melanoma cell growth and progression.
Main Methods:
- Cell viability was assessed using MTT assays.
- Cell cycle, apoptosis, migration, invasion, and mitochondrial function were evaluated using flow cytometry, Western blotting, and specific assays.
- The PI3K/AKT/p21 pathway was analyzed.
Main Results:
- Methanol extract of immature P. trifoliata (MEPT) significantly inhibited B16 cell viability, migration, and invasion in a dose- and time-dependent manner.
- MEPT induced G1 cell cycle arrest and apoptosis by modulating Bax, Apaf-1, and Bcl-2 expression.
- The extract disrupted mitochondrial membrane potential and activated the PI3K/AKT/p21 pathway.
Conclusions:
- Immature P. trifoliata extract demonstrates anti-melanoma activity by inducing apoptosis through mitochondrial pathways.
- The findings suggest that MEPT is a promising candidate for further development as a melanoma therapeutic agent.
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