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Anti-proliferative effect of Annona extracts on breast cancer cells
Maria-Luisa Veisaga1,2, Mariam Ahumada2, Stacy Soriano2
1Biomolecular Sciences Institute, Florida International University, Miami, 33199, USA.
Backgorund:
Fruits and seed extracts of Annona montana have significant cytotoxic potential in several cancer cells. This study evaluates the effect of A. montana leaves hexane extract on several signaling cascades and gene expression in metastatic breast cancer cells upon insulin-like growth factor-1 (IGF-1) stimulation.
Methods:
MTT assay was performed to determine the proliferation of cancer cells. Propidium iodide staining and flow cytometry analysis of Annexin V binding was utilized to measure the progression of the cell cycle and the induction of apoptosis. Protein expression and phosphorylation were determined by western blotting analysis to examine the underlying cellular mechanism triggered upon treatment with A. montana leaves hexane extract.
Results:
A. montana leaves hexane (sub-fraction V) blocked the constitutive stimulation of the PI3K/mTOR signaling pathways. This inhibitory effect was associated with apoptosis induction as evidenced by the positivity with Annexin V and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNNEL) staining, activation of caspase-3, and cleavage of PPAR. It also limited the expression of various downstream genes that regulate proliferation, survival, metastasis, and angiogenesis (i.e., cyclin D1, survivin, COX-2, and VEGF). It increased the expression of p53 and p21. Interestingly, we also observed that this extract blocked the activation of AKT and ERK without affecting the phosphorylation of the IGF-1 receptor and activation of Ras upon IGF-1 stimulation.
Conclusion:
Our study indicates that A. montana leaves (sub-fraction V) extract exhibits a selective anti-proliferative and proapoptotic effect on the metastatic MDA-MB-231 breast cancer cells through the involvement of PI3K/AKT/mTOR/S6K1 pathways.
Insights
Annona montana leaf extract selectively inhibits metastatic breast cancer cell growth and promotes apoptosis. It targets key signaling pathways like PI3K/AKT/mTOR, offering a potential therapeutic avenue.
Area of Science:
- Molecular Biology
- Cancer Research
- Pharmacology
Background:
- Annona montana extracts show cytotoxic effects against various cancer cells.
- This study investigates the impact of A. montana leaf hexane extract on signaling pathways and gene expression in metastatic breast cancer cells.
Purpose of the Study:
- To evaluate the effect of Annona montana leaves hexane extract on signaling cascades in metastatic breast cancer cells.
- To analyze gene expression changes induced by the extract upon insulin-like growth factor-1 (IGF-1) stimulation.
Main Methods:
- MTT assay for cell proliferation assessment.
- Flow cytometry with Annexin V and propidium iodide staining for cell cycle analysis and apoptosis.
- Western blotting to determine protein expression and phosphorylation levels.
Main Results:
- A. montana leaf hexane extract (sub-fraction V) inhibited PI3K/mTOR signaling pathways.
- The extract induced apoptosis, evidenced by Annexin V/TUNNEL staining, caspase-3 activation, and PPAR cleavage.
- Gene expression of cyclin D1, survivin, COX-2, and VEGF was reduced, while p53 and p21 expression increased. AKT and ERK activation was blocked without affecting IGF-1 receptor phosphorylation.
Conclusions:
- Annona montana leaf extract (sub-fraction V) demonstrates selective anti-proliferative and proapoptotic effects on metastatic MDA-MB-231 breast cancer cells.
- The observed effects are mediated through the PI3K/AKT/mTOR/S6K1 signaling pathways.
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