MON-266 The Effects of Azadirachta Indica (Neem) on the Expression of Estrogen Receptor and Tumor Suppressor Genes (BRCA1 and P53) in Breast Cancer Cells

Abstract Disclosure: J.J. Koujane: None. N. Prudhomme: None. R.R. Yaldo: None. A. Zanib: None. S.E. Pfiffner: None. I. Iskander: None. E. Lumaj: None. S. Dinda: None. Naturally sourced compounds are increasingly sought after for disease treatment, including breast cancer. Azadirachta Indica, commonly known as Neem, has a long-standing history of use in traditional Chinese and Indian medicine, and it has gained significant attention in modern research due to its therapeutic properties. Recent studies have begun to explore Neem’s potential as an anti-inflammatory agent, particularly against breast cancer, the most frequently diagnosed cancer and the leading cause of cancer-related deaths among women. Despite its promising therapeutic potential, there are still gaps in understanding the biochemical mechanisms and pathways underlying Neem’s beneficial properties. Approximately 55-65% of women with the BRCA1 gene mutation will develop hereditary breast cancer; therefore, exploring treatment options for receptor-positive patients is critical. This study examines the effects of Neem, alone and in combination with hormones and anti-hormones, on estrogen receptor alpha (ERα), p53, and BRCA1 expression in MCF-7 and T-47D breast cancer cells utilizing western blot analysis, cellular viability assays, confocal microscopy, and RT-qPCR analyses. To ensure that treatment conditions are absent of endogenous steroids or growth factors, cells were cultured in a medium containing 5% dextran-coated charcoal-treated fetal bovine serum (DCC-FBS) for 6 days. For western blot analyses, cells were treated with various concentrations of Neem ranging from 10-100 µM for 24 hours. Compared to the control, a concentration-dependent decrease of ERα, BRCA1, and p53 expression was noted in both cell lines. In our hormone studies, we utilized the optimal concentration of Neem (80 μM) alone and in combination with hormones and anti-hormones. After 24-hour treatment of Neem, 17-β-estradiol (E2), and E2 with Neem, we observed a decrease in ERα and BRCA1 expression compared to the control. Combination treatment of Neem with anti-hormone ICI revealed a significant decrease in ERα, BRCA1, in T-47D and MCF-7 cells. The same treatment conditions in MCF-7 cells revealed a significant decrease in expression of ERα and BRCA1 when compared to the control. Image cytoNmetric analysis with propidium iodide staining was performed to examine Neem’s effects on cellular viability in T-47D and MCF-7 cells. Cells treated for 6 days with 80 μM Neem in combination with hormone and anti-hormones displayed a decrease in cell proliferation compared to the control in both cell lines. Cytolocalization studies revealed no significant change in ERα location after treatment with Neem. RT-qPCR experiments are currently in progress to further elucidate the effects of Neem on the transcriptional profile of ESR1 and BRCA1 genes. Our research offers a promising direction for exploring the molecular mechanisms of Neem in regulating steroid receptors and tumor suppressor genes in breast cancer cells. Presentation: Monday, July 14, 2025