Dual-Method Apoptosis Evaluation Reveals the Therapeutic Potential of Garcinia cowa Extract in Combination with Doxorubicin for Breast Cancer Treatment

Breast cancer remains a major health challenge worldwide, demanding the exploration of novel therapeutic agents with improved efficacy and safety. Garcinia cowa, a traditional medicinal plant, has shown promising cytotoxic activity in previous studies. This study aimed to evaluate the apoptosis-inducing potential of Garcinia cowa stem bark ethanol extract (GCSBEE) at 130 µg/mL, either alone or in combination with doxorubicin (Doxo) (0.026 µg/mL), in human breast cancer T47D cells. T47D breast cancer cells were treated with GCSBEE at 130 µg/mL, doxorubicin at 0.026 µg/mL, and their combination (GCSBEE + Doxo) for 48 h. After treatment, Apoptosis was assessed using dual staining with acridine orange/propidium iodide and flow cytometry analysis. The results demonstrated that both the extract and doxorubicin significantly increased the proportion of apoptotic cells in both methods. Co-treatment with GCSBEE and Doxo demonstrated a marked shift in cell death dynamics, favoring apoptosis over necrosis. AO/PI staining indicated a significantly elevated apoptotic rate alongside minimal necrotic activity. These results were consistently supported by flow cytometry data, which confirmed a higher proportion of early and late apoptotic cells compared to necrotic cells. Collectively, these findings indicate that GCSBEE may play a modulatory role in enhancing the apoptotic response induced by Doxorubicin, potentially contributing to a more targeted and controlled form of cell death.