Modulation of BAX/BCL-2 Expression and Cyclophilin A-Targeted In Silico Study by Spatholobus littoralis Combined with Doxorubicin in DMBA-Induced Breast Cancer Rat Model
DOI:
https://doi.org/10.48048/tis.2026.13669Keywords:
BAX/BCL-2 expression ratio, Cyclophilin A, DMBA-induced breast cancer, Doxorubicin, Molecular docking, Spatholobus littoralisAbstract
Cyclophilin A (CypA) has been identified as a key regulator of apoptosis and contributes to chemotherapy resistance in breast cancer cells. One indicator of apoptosis imbalance is the BAX/BCL-2 ratio, where a low ratio is associated with tumor progression and reduced treatment efficacy. Overcoming this resistance remains a challenge, particularly through combination therapies. Spatholobus littoralis, a flavonoid-rich medicinal plant, has shown potential as an anticancer agent, but its effectiveness as an adjuvant to doxorubicin is not fully established. This study aims to evaluate the effects of S. littoralis extract combined with doxorubicin on apoptosis regulation, particularly the BAX/BCL-2 expression ratio, in a DMBA-induced breast cancer rat model, and to investigate the interaction of its major compound with Cyclophilin A through molecular docking analysis. Female Sprague Dawley rats were induced with DMBA to develop breast tumors and subsequently treated with S. littoralis extract, doxorubicin, or their combination. Gene expression levels of BAX and BCL-2 were quantified using real-time PCR, and tumor volume and BW were monitored weekly. Data were analyzed using one-way ANOVA with post hoc tests (p < 0.05). Molecular docking evaluated the interaction between N-2-[4-(3,3-dimethylallyloxy)phenyl]ethylcinnamide, the plant’s main compound, and CypA (PDB ID: 3RDD), while SwissADME assessed drug-likeness and pharmacokinetics. Results showed that DMBA increased BCL-2 and decreased BAX, indicating apoptosis resistance. Treatment with S. littoralis extract combine with doxorubicin individually significantly improved the BAX/BCL-2 ratio, reduced tumor volume, and increased %TGI, with the combination therapy producing the most pronounced pro-apoptotic effect (p < 0.05) and maintaining BW. Molecular docking revealed strong binding affinity of the compound to CypA, and in silico analysis suggested favorable drug-like properties. In conclusion, the combination of S. littoralis extract with doxorubicin enhances breast cancer treatment efficacy by modulating apoptosis markers, reducing tumor growth, and targeting Cyclophilin A, while maintaining body weight.
HIGHLIGHTS
- Combination therapy using doxorubicin and Spatholobus extract inhibited tumor growth in DMBA-induced breast cancer rats
- Combination treatment enhanced intrinsic apoptotic signaling through modulation of the BAX/BCL-2 ratio
- Combination therapy demonstrated stronger pro-apoptotic responses compared with doxorubicin monotherapy
- Bioactive compounds from Spatholobus extract exhibited strong binding affinity toward Cyclophilin A (CypA) in molecular docking analysis
- Integrated in vivo and in silico findings suggested the involvement of CypA-related apoptotic pathways in breast cancer suppression
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