Inhibition of EGF and CoCl2-Induced HIF-1α and VEGF Production in Triple Negative MDA-MB-468 Cells by Umbelliprenin: Unveiling the Role of PI3K/AKT/mTOR and MAPK Pathways

Background and objectives: Triple-negative breast cancer is a significant global health challenge, and there's growing interest in targeting multiple pathways for treatment. Umbelliprenin, derived from herbal sources, has shown anti-tumor potential. This study aimed to assess umbelliprenin's impact on key genes related to proliferation, metastasis, and angiogenesis. Methods: Umbelliprenin, which was synthesized by the Pharmaceutical Research Center (PRC) at Mashhad University of Medical Sciences in Iran, was utilized in this study. The study aimed to investigate the impact of umbelliprenin on EGF and CoCl2-induced signaling in the PI3K/AKT/mTOR and MAPK pathways. Quantitative PCR was employed to assess the expression of EGFR, PI3K, AKT, mTOR, S6K, ERK1, ERK2, 4EBP1, HIF-1α, HIF-1β, VEGF, and VEGFR genes. Additionally, immunoblot assays were conducted to evaluate the levels of VEGF and HIF-1α in MDA-MB-468 cells. Results: The study found that umbelliprenin had cytotoxic effects, with an IC50 value of 152.5 µM. At concentrations of 10 µM and 20 µM, it upregulated genes like EGFR, VEGFR, HIF-1α, VEGF, PI3K, ERK2, and mTOR while downregulating 4EBP1. Umbelliprenin also increased VEGF protein levels. When used on EGF-stimulated cells, it enhanced VEGF and PI3K expression while inhibiting AKT, ERK2, mTOR, and antiproliferative 4EBP1 genes. Notably, VEGF and HIF-1α protein levels remained unchanged. Conversely, umbelliprenin downregulated EGFR, AKT, ERK1/2, HIF-1α, and VEGF in CoCl2-stimulated cells, while elevating 4EBP1 and reducing VEGF and HIF-1α protein levels. Conclusion: Umbelliprenin inhibited MDA-MB-468 cell growth and impacted gene expression related to metastasis and angiogenesis, particularly under conditions of EGFR activation and hypoxia