PJB-2024-66
ACHILLEA FRAGRANTISSIMA (FORSSK.) SCH.BIP INSTIGATES THE ROS/FADD/C-PARP EXPRESSION: AN ACTUATION OF APOPTOSIS IN BREAST CANCER CELL
Abdulrahman Alasmari
Abstract
Achillea fragrantissima (A. fragrantissima) is a shrub that belongs to the Asteraceae family in Arabia and Egypt. It is used as a folk medicine and is a strong source of phenolic acids, flavonoids, and some active compounds. Here, we investigated the anti-cancer effect of A. fragrantissima on breast cancer MCF-7 cells and the critical mechanism involved in apoptosis. The ethanolic extract of A. fragrantissima was examined for anti-breast cancer properties. In turn, cytotoxicity (IC50 value) and cell viability were assessed using the MTT method. Additionally, trypan blue exclusion and microscopy examination demonstrated the presence of apoptotic cells. Fluorescent staining, such as of AO/EtBr, DCFH-DA, Rho-123, and Hoechst-33342, revealed the cellular cytoplasmic disciplines upon A. fragrantissima effect. Moreover, cellular functioning tests, such as wound healing, colony formation, and a transwell invasion, assay were performed. The qRT-PCR technique was used to authenticate the A. fragrantissima-induced apoptotic network genes (Caspase-3, Caspase-8, Caspase-9, Cytochrome c, BCL-2, BID, BAX, PARP, PTEN, PI3K, and Akt) and expression was evaluated. The immunoblot technique was used to assess the expressed levels of apoptotic proteins, such as cleaved PARP, CYSC, and FADD. Thus, this study found that A. fragrantissima exerted cytotoxicity at 20 µg/mL for 24 hours in MCF-7 cells. We also found that it decreased cellular viability, produced apoptotic cells, and damaged cellular surfaces with dead matter. Consequently, this created ROS species accumulation, loss of mitochondrial membrane potential, and fragmentation of DNA in MCF-7 cells. Furthermore, it arrested cell migration, induced colony-forming ability loss, and suppressed cell invasion. A. fragrantissima also significantly upregulated genes such as caspase-3, 9, cytochrome c, BID, BAX, and PTEN, while downregulating the Pi3K/ Akt signaling. Nonetheless, A. fragrantissima induced cleaved PARP, CYSC, and FADD proteins in MCF-7 cells to avail apoptosis.
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