UPLC-Q-TOF/MS-Based Metabolomics Approach Reveals Osthole Intervention in Breast Cancer 4T1 Cells
Xiuyun Li1, Chenglun Zhang1, Enhui Wu2
1School of Pharmacy, The Center for Drug Research and Development, Guangdong Pharmaceutical University, Guangzhou 510006, China.
Abstract:
Osthole (OST) is a simple coumarin derivative with pharmacological effects in many types of cancer cells. However, its role and its mechanism of action in breast cancer 4T1 cells remain unclear. In this study, we explored the effects and potential mechanisms of action of OST in 4T1 cells. The MTT, PI, and Annexin V-FITC/PI methods were used to evaluate the effects of OST-treated and untreated 4T1 cells on viability, cell cycle, and apoptosis, respectively. UPLC-Q-TOF/MS combined with multivariate data analysis was used to screen potential biomarkers relevant to the therapeutic mechanisms of OST. Additionally, mTOR, SREBP1, and FASN protein levels were detected using western blotting in OST-treated and untreated 4T1 cells. OST inhibited 4T1 cell proliferation, blocked the cells from remaining in S-phase, and induced apoptosis. In 4T1 cells, OST mainly affected the phospholipid biosynthesis, methyl histidine metabolism, pyrimidine metabolism, and β-oxidation of very long chain fatty acid pathways, suggesting that metabolic changes related to lipid metabolism-mediated signaling systems were the most influential pathways, possibly via inhibition of mTOR/SREBP1/FASN signaling. Our findings reveal biomarkers with potential therapeutic effects in breast cancer and provide insight into the therapeutic and metabolic mechanisms of OST in 4T1 cells.
Insights
Osthole (OST) inhibits breast cancer 4T1 cell proliferation and induces apoptosis. This study reveals OST
Area of Science:
- Pharmacology and Molecular Biology
- Cancer Research
- Metabolomics
Background:
- Osthole (OST), a coumarin derivative, exhibits anticancer properties.
- The specific mechanisms of OST action in 4T1 breast cancer cells are not well understood.
- Investigating OST's effects is crucial for developing novel breast cancer therapies.
Purpose of the Study:
- To elucidate the effects of OST on 4T1 breast cancer cells.
- To identify the underlying molecular and metabolic mechanisms of OST action.
- To explore OST as a potential therapeutic agent for breast cancer.
Main Methods:
- Cell viability assessed using MTT assay.
- Cell cycle analysis performed with Propidium Iodide (PI) staining.
- Apoptosis detection via Annexin V-FITC/PI staining.
- Metabolomic profiling using UPLC-Q-TOF/MS and multivariate data analysis.
- Protein expression analysis of mTOR, SREBP1, and FASN via Western blotting.
Main Results:
- OST significantly inhibited 4T1 cell proliferation and induced apoptosis.
- OST treatment led to cell cycle arrest in the S-phase.
- Metabolomic analysis identified alterations in phospholipid biosynthesis, methyl histidine metabolism, pyrimidine metabolism, and fatty acid oxidation pathways.
- OST suppressed the expression of mTOR, SREBP1, and FASN proteins.
- These findings suggest a mechanism involving the inhibition of the mTOR/SREBP1/FASN signaling pathway.
Conclusions:
- Osthole demonstrates potent anti-proliferative and pro-apoptotic effects on 4T1 breast cancer cells.
- OST's therapeutic action is associated with significant metabolic reprogramming, particularly in lipid metabolism.
- The study identifies OST as a promising candidate for breast cancer treatment, highlighting its impact on key signaling pathways and metabolic processes.
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