Combination of Eleutherine bulbosa Extract and Tamoxifen  Suppresses 7,12-dimethylbenz[a]anthracene (DMBA)-Induced  Breast Cancer: In Vivo Evidence of Enhanced p53 Expression  and Histopathological Improvement

Fifin Ernawati; Andi Nilawati Usman; Risfah Yulianty; Andi Ariyandy

doi:10.31557/apjcb.2026.11.1.109-118

Authors

Fifin Ernawati Department of Midwifery, Graduate School, Hasanuddin University, Makassar, South Sulawesi, Indonesia. https://orcid.org/0009-0004-5326-3848
Andi Nilawati Usman Department of Midwifery, Graduate School, Hasanuddin University, Makassar, South Sulawesi, Indonesia. https://orcid.org/0000-0002-1136-1704
Risfah Yulianty Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Hasanuddin University, Makassar, South Sulawesi, Indonesia. https://orcid.org/0009-0009-8127-5084
Andi Ariyandy Department of Physiology, Faculty of Medicine, Hasanuddin University, Makassar, South Sulawesi, Indonesia. https://orcid.org/0000-0002-6492-5235

DOI:

https://doi.org/10.31557/apjcb.2026.11.1.109-118

Keywords:

Eleutherine bulbosa, Tamoxifen, Breast Neoplasms, Tumor Suppressor Protein p53, Integrative Medicine

Abstract

Background: Eleutherine bulbosa (Dayak onion), a phytotherapeutic plant rich in flavonoids and polyphenols, exhibits potent anticancer activity. As monotherapy frequently leads to treatment resistance, combining Eleutherine bulbosa ethanolic extract with standard endocrine therapy such as tamoxifen offers a promising approach to overcome resistance and improve treatment outcomes in breast cancer. This study evaluated whether simultaneous administration improves p53 activation and restores mammary tissue architecture in a DMBA-induced breast cancer mouse model.

Materials and Methods: An in vivo post-test-only design was employed, involving 36 female BALB/c mice (n = 6 per group), including monotherapy, sequential combination (I3), and simultaneous combination (I4) regimens. Primary endpoints included histopathological evaluation of mammary tissue and quantification of p53 protein using Enzyme-Linked Immunosorbent Assay (ELISA) and Immunohistochemistry (IHC). Histological architecture was assessed by Hematoxylin and Eosin (HE) staining. Histology was performed on a single representative sample per group (n = 1), therefore interpreted descriptively.

Results: In vivo findings demonstrated that the simultaneous combination (I4) provided the most comprehensive therapeutic benefit. Group I4 showed the highest plasma p53 level (8.70 ± 0.30 pg/mL), which was significantly higher than that of the positive control (KP) but statistically comparable to tamoxifen monotherapy (I2, 8.09 ± 0.40 pg/mL). Similarly, the percentage of p53-positive cells detected by IHC did not differ significantly between I4 and I2. However, group I4 displayed the most pronounced morphological recovery, exhibiting the highest mean alveolar diameter (106.71 µm) and the lowest mean alveolar density (1.8/field), indicating the most favorable histological trend in reversing tissue damage compared to I2.

Conclusion: Simultaneous administration of Eleutherine bulbosa extract with tamoxifen provided the most favorable morphological trend compared with monotherapies, likely through localized, tissue-level actions rather than systemic p53 elevation.