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In Vitro Analysis of Cytotoxic Activities of <i>Monotheca buxifolia</i> Targeting <i>WNT</i>/<i>β-Catenin</i> Genes in Breast Cancer Cells
oleh: Ambreen Sher, Sobia Tabassum, Heather Mann Wallace, Asifullah Khan, Asad Mustafa Karim, Sarah Gul, Se Chan Kang
| Format: | Article |
|---|---|
| Diterbitkan: | MDPI AG 2023-03-01 |
Deskripsi
Breast cancer (BC) is known to be the most common malignancy among women throughout the world. Plant-derived natural products have been recognized as a great source of anticancer drugs. In this study, the efficacy and anticancer potential of the methanolic extract of <i>Monotheca buxifolia</i> leaves using human breast cancer cells targeting <i>WNT</i>/<i>β-catenin</i> signaling was evaluated. We used methanolic and other (chloroform, ethyl acetate, butanol, and aqueous) extracts to discover their potential cytotoxicity on breast cancer cells (MCF-7). Among these, the methanol showed significant activity in the inhibition of the proliferation of cancer cells because of the presence of bioactive compounds, including phenols and flavonoids, detected by a Fourier transform infrared spectrophotometer and by gas chromatography mass spectrometry. The cytotoxic effect of the plant extract on the MCF-7 cells was examined by MTT and acid phosphatase assays. Real-time PCR analysis was performed to measure the mRNA expression of <i>WNT-3a</i> and <i>β-catenin</i>, along with <i>Caspase-1,-3,-7,</i> and <i>-9</i> in MCF-7 cells. The IC50 value of the extract was found to be 232 μg/mL and 173 μg/mL in the MTT and acid phosphatase assays, respectively. Dose selection (100 and 300 μg/mL) was performed for real-time PCR, Annexin V/PI analysis, and Western blotting using Doxorubicin as a positive control. The extract at 100 μg/mL significantly upregulated caspases and downregulated the <i>WNT-3a</i> and <i>β-catenin</i> gene in MCF-7 cells. Western blot analysis further confirmed the dysregulations of the <i>WNT</i> signaling component (*** <i>p<</i> 0.0001). The results showed an increase in the number of dead cells in methanolic extract-treated cells in the Annexin V/PI analysis. Our study concludes that <i>M. buxifolia</i> may serve as an effective anticancer mediator through gene modulation that targets <i>WNT</i>/<i>β-catenin</i> signaling, and it can be further characterized using more powerful experimental and computational tools.