The Synergistic and Anticancer Potential of Withania Somnifera (Ashwagandha) Ethanol Extract as an Adjuvant with Doxorubicin in MCF7 Breast Cancer Cell Line.

BACKGROUND: Recent approaches in breast cancer treatment involve exploring natural extracts either as substitutes for traditional chemotherapeutic agents or as adjuvants to enhance their effectiveness. In this context, ethanolic extracts from Withania Somnifera (WS) and Arctium Lappa (AL) have gained attention for their potential anticancer properties across various cancer types. OBJECTIVE: This study aimed to investigate the in vitro anticancer efficacy of ethanolic extracts from both WS and AL in the MCF7 breast cancer cell line. METHODS: The study encompassed assessments of total antioxidant activity, total flavonoids, total phenolic compounds, and cytotoxicity for each extract. Unraveling the molecular mechanisms underlying the action of these extracts involved Real-time PCR and flow cytometry to analyze apoptotic markers and cell cycle dynamics, respectively. RESULTS: Results indicated that the WS ethanolic extract exhibited better efficacy, especially in terms of IC50. To explore its potential as an adjuvant with doxorubicin (DOX) in breast cancer therapy, 16 non-constant combination ratios were tested. The combination of 1/2 WS to 1/4 DOX, with a combination index (CI) of -0.229 was selected for further downstream molecular analysis. Gene expression analysis revealed a significant upregulation (p < 0.05) of P53, Cas3, Cas7, and Bax. Flow cytometric analysis further highlighted cell cycle arrest at the S phase. Additionally, the study explored the synergistic inhibitory effects of WS and DOX on key enzymes in the glycolytic pathway, namely, pyruvate kinase and aldolase. CONCLUSION: In conclusion, the ethanolic extract of WS demonstrated a synergistic effect as an adjuvant with DOX, showing promise as a combination therapy for breast cancer.