The Impact of Amphimedon chloros Ethyl Acetate Extract on MDA-MB-231 Cell Lines' Expression of Bax, Cyclin D, Caspase 3, P21, C-myc, and Bcl2: Mechanism of inhibition of the Growth of Cancer Cells

Numerous marine sponge bioactive compounds exhibit diverse pharmacological properties. This study investigated the ethyl acetate extract of Amphimedon chloros for its effects on breast cancer cells (MDA-MB-231), focusing on the expression of key genes involved in apoptosis and cell cycle regulation, including Bax, Cyclin D, caspase 3, p21, C-myc, and Bcl2. The extract's chemical profile was analyzed using Liquid Chromatography-Mass Spectrometry (LC-MS), identifying fourteen compounds such as zamamidine (11.4%), keramamine (10.2%), and methoxyhexadecanoate (9.1%). Cytotoxicity and anticancer activity were assessed on MDA-MB-231 cells and human periodontal ligament fibroblasts (HPLFs). The IC50 of the extract against MDA-MB-231 cells was 3.0 µg/mL, indicating significant cytotoxicity, while it did not affect fibroblast viability at concentrations ranging from 50-200 µg/mL. RT-PCR (Real Time-Polymerase Chain Reaction) analysis revealed that the extract increased the expression of Bax, p21, and caspase 3, and decreased the levels of Bcl2 and Cyclin D in MDA-MB-231 cells. These findings suggest that the extract may inhibit cancer cell proliferation by interfering with growth factors signaling pathways, such as PI3K/Akt (Phosphoinositide 3-Kinase/Protein Kinase B) and MAPK (Mitogen-Activated Protein Kinase). The increase in Bax and p21 expression indicates enhanced apoptosis, while the drastic reduction in c-Myc expression points to potential cell cycle arrest. The results support the potential of A. chloros as an anticancer agent, with selective cytotoxicity towards cancer cells and significant modulation of apoptotic and cell cycle-related genes.