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Decursinol Angelate Arrest Melanoma Cell Proliferation by Initiating Cell Death and Tumor Shrinkage via Induction of
Sukkum Ngullie Chang1,2, Imran Khan2,3, Chang Geon Kim1
1Advanced Bio Convergence Center (ABCC), Pohang Technopark Foundation, Pohang, Gyeongbuk 37668, Korea.
Abstract:
Melanoma is known to aggressively metastasize and is one of the prominent causes of skin cancer mortality. This study was designed to assess the molecular mechanism of decursinol angelate (DA) against murine melanoma cell line (B16F10 cells). Treatment of DA resulted in growth inhibition and cell cycle arrest at G0/G1 (p < 0.001) phase, evaluated through immunoblotting. Moreover, autophagy-related proteins such as ATG-5 (p < 0.0001), ATG-7 (p < 0.0001), beclin-1 (p < 0.0001) and transition of LC3-I to LC3-II (p < 0.0001) were markedly decreased, indicating autophagosome inhibition. Additionally, DA treatment triggered apoptotic events which were corroborated by the occurrence of distorted nuclei, elevated reactive oxygen species (ROS) levels and reduction in the mitochondrial membrane potential. Subsequently, there was an increase in the expression of pro-apoptotic protein Bax in a dose-dependent manner, with the corresponding downregulation of Bcl-2 expression and cytochrome C expression following 24 h DA treatment in A375.SM and B16F10 cells. We substantiated our results for apoptotic occurrence through flow cytometry in B16F10 cells. Furthermore, we treated B16F10 cells with N-acetyl-L-cysteine (NAC). NAC treatment upregulated ATG-5 (p < 0.0001), beclin-1 (p < 0.0001) and LC3-I to LC3-II (p < 0.0001) conversion, which was inhibited in the DA treatment group. We also noticed a systematic upregulation of important markers for progression of G1 cell phase such as CDK-2 (p < 0.029), CDK-4 (p < 0.036), cyclin D1 (p < 0.0003) and cyclin E (p < 0.020) upon NAC treatment. In addition, we also observed a significant fold reduction (p < 0.05) in ROS fluorescent intensity and the expression of Bax (p < 0.0001), cytochrome C (p < 0.0001), cleaved caspase-9 (p > 0.010) and cleaved caspase-3 (p < 0.0001). NAC treatment was able to ameliorate DA-induced apoptosis and cell cycle arrest to support our finding. Our in vivo xenograft model also revealed similar findings, such as downregulation of CDK-2 (p < 0.0001) and CDK-4 (p < 0.0142) and upregulation of Bax (p < 0.0001), cytochrome C (p < 0.0001), cleaved caspase 3 (p < 0.0001) and cleaved caspase 9 (p < 0.0001). In summary, our study revealed that DA is an effective treatment against B16F10 melanoma cells and xenograft mice model.
Insights
Decursinol angelate (DA) effectively inhibits melanoma cell growth by inducing cell cycle arrest and apoptosis. This compound also suppresses autophagy and reduces reactive oxygen species (ROS), offering a promising therapeutic strategy for melanoma treatment.
Area of Science:
- Oncology
- Pharmacology
- Molecular Biology
Background:
- Melanoma is a significant cause of skin cancer mortality due to its aggressive metastatic potential.
- Understanding the molecular mechanisms underlying melanoma progression is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the molecular mechanisms of decursinol angelate (DA) in inhibiting murine melanoma cell line (B16F10) growth.
- To assess the effects of DA on cell cycle, autophagy, apoptosis, and reactive oxygen species (ROS) in melanoma cells.
Main Methods:
- Treatment of B16F10 melanoma cells with DA.
- Evaluation of cell cycle progression using immunoblotting.
- Assessment of autophagy markers (ATG-5, ATG-7, beclin-1, LC3-I/II).
- Analysis of apoptotic events including nuclear morphology, ROS levels, mitochondrial membrane potential, and expression of apoptosis-related proteins (Bax, Bcl-2, cytochrome C).
- In vitro and in vivo xenograft models were utilized.
Main Results:
- DA treatment induced significant G0/G1 cell cycle arrest (p < 0.001) and inhibited autophagy by decreasing key proteins (ATG-5, ATG-7, beclin-1, LC3-II).
- DA triggered apoptosis, evidenced by distorted nuclei, increased ROS, reduced mitochondrial membrane potential, and altered Bax/Bcl-2/cytochrome C expression.
- N-acetyl-L-cysteine (NAC) treatment reversed DA-induced effects, confirming the role of ROS and validating DA's mechanism.
- In vivo studies corroborated in vitro findings, showing DA's efficacy in inhibiting tumor growth and promoting apoptosis.
Conclusions:
- Decursinol angelate (DA) demonstrates significant anti-melanoma activity by inhibiting cell proliferation, inducing apoptosis, and suppressing autophagy.
- DA's mechanism involves the modulation of cell cycle regulators, apoptotic pathways, and ROS levels.
- DA represents a potential therapeutic agent for melanoma treatment, warranting further investigation.
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