Amphotericin-A21 produces antiproliferative effects through modulating EGFR expression in glioblastoma multiforme cells

• Published in: The Journal of pharmacology and experimental therapeutics 2024-06, Vol.389, p.59-59
• Main Authors: Rodriguez-Fragoso, Lourdes, Zamora-Morán, Esdras, Rodríguez-López, Anahi, Ortega-Blake, Ivan, Hernandez, Arturo Galvan, Diaz-Peralta, Lucero
• Format: Article
• Language: English
• ISSN: 0022-3565
• DOI: 10.1124/jpet.059.131002

Abstract ID 131002 Poster Board 059 Glioblastoma multiforme (GBM) is the most frequent malignant astrocytic tumor of the central nervous system (CNS). The survival time of patients with GBM is 8 to 15 months. Our research group has developed a semi-synthetic compound derived from amphotericin B, called Amphotericin-A21 (AmB-A21), which has shown to have a better biosafety profile in animal models and in non-tumor cells (HEK-293) compared to its precursor. Recently, AmB-A21 was shown to induce cytotoxicity in breast cancer cell lines (MDA-MB-231 and MCF-7). In addition, AmB-A21 has been shown to have a synergistic effect with carboplatin inducing apoptosis and inhibiting proliferation and cell invasion of lung adenocarcinoma cells (A549). The aim of this work was to determine the effect of AmB-A21 on EGFR and it signaling pathways involved in tumor proliferation and invasion in GBM cells. Cell viability and cell proliferation were determined using MTS (CellTiter 96® AQueous One Solution de Promega). Briefly, for cell viability, U-87 MG (HTB-14 TM; ATCC) cells were seeded into a 96-well plate (10,000/well) and incubated for 24 h; for cell proliferation cells were incubated during 24, 48 and 72 h at 37°C and 5% CO2. The culture medium was replaced by a fresh one supplemented with different concentrations of AmB-A21 (1 to 300 μM) and incubated for 24 h. After treatment, MTS-PMS was added and incubated for 1 h. The optical density was determined on a microplate reader (Bio-Rad) at a wavelength of 490 nm and we calculated inhibitory concentration 25 (IC25) and the effect on proliferation rates. Morphological analysis was performed by using hemotoxylin and eosin (H&E) staining of U-87 MG cells. Cells were seeded into a 24-well plate (50,000/well) and treated for 24 h with AmB-A21, as described above. Finally, microphotographs were taken in a light microscopy (Olympus Microscope). We evaluated cell migration with the wound-healing assay after 24h of treatment. Microphotographs were taken and migrating cells were counted. We analyze the expression of phosphosite pY1173-EGFR and PLCγ1 by immunocytochemistry. AmB-A21 had cytotoxic effect and inhibited the proliferation of U-87MG cells in a concentration- and time-dependent manner, respectively (p