Red Sea Sponge Callyspongia siphonella Extract Induced Growth Inhibition and Apoptosis in Breast MCF-7 and Hepatic HepG-2 Cancer Cell Lines in 2D and 3D Cell Cultures

Abstract

Sana A Fadil,1,2 Fadwa A Aljoud,2,3 Ahmed R Yonbawi,1 Ahmad J Almalki,4 Rawan H Hareeri,5 Abrar Ashi,2,6 Mehal Atallah AlQriqri,2 Nada S Bawazir,2 Hadeel H Alshangiti,2 Lamiaa A Shaala,7,8 Diaa TA Youssef,1,8,9 Faris A Alkhilaiwi1,2 1Department of Natural Products and Alternative Medicine, Faculty of Pharmacy, King Abdulaziz University, Jeddah, 21589, Saudi Arabia; 2Regenerative Medicine Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, 21589, Saudi Arabia; 3Scientific Research Center, Dar Al-Hekma University, Jeddah, 22246, Saudi Arabia; 4Department of Pharmaceutical Chemistry, Faculty of Pharmacy, King Abdulaziz University, Jeddah, 21589, Saudi Arabia; 5Department of Pharmacology and Toxicology, Faculty of Pharmacy, King Abdulaziz University, Jeddah, 21589, Saudi Arabia; 6Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, 21589, Saudi Arabia; 7Suez Canal University Hospital, Suez Canal University, Ismailia, 41522, Egypt; 8Natural Products Unit, King Fahad Medical Research Center, King Abdulaziz University, Jeddah, 21589, Saudi Arabia; 9Department of Pharmacognosy, Faculty of Pharmacy, Suez Canal University, Ismailia, 41523, EgyptCorrespondence: Sana A Fadil; Faris A Alkhilaiwi, Email [email protected]; [email protected]: The increasing incidence of cancer diseases necessitates the urgent exploration of new bioactive compounds. One of the trends in drug discovery is marine sponges which is gaining significant support due to the abundant production of natural pharmaceutical compounds obtained from marine ecosystems. This study evaluates the anticancer properties of an organic extract from the Red Sea sponge Callyspongia siphonella (C. siphonella) on HepG-2 and MCF-7 cancer cell lines.Methods: C. siphonella was collected, freeze-dried, and extracted using a methanol-dichloromethane mixture. The extract was analyzed via Liquid Chromatography-Mass Spectrometry. Cytotoxic effects were assessed through cell viability assays, apoptosis detection, cell cycle analysis, mitochondrial membrane potential assays, scratch-wound healing assays, and 3D cell culture assays.Results: Fifteen compounds were identified in the C. siphonella extract. The extract showed moderate cytotoxicity against MCF-7 and HepG-2 cells, with IC50 values of 35.6 ± 6.9 μg/mL and 64.4 ± 8 μg/mL, respectively, after 48 hours of treatment. It induced cell cycle arrest at the G2/M phase in MCF-7 cells and the S phase in HepG-2 cells. Apoptosis increased significantly in both cell lines, accompanied by reduced mitochondrial membrane potential. The extract inhibited cell migration, with notable reductions after 24 and 48 hours. In 3D cell cultures, the extract had IC50 values of 5.1 ± 2 μg/mL for MCF-7 and 166.4 ± 27 μg/mL for HepG-2 after 7 days of treatment, showing greater potency in MCF-7 spheres compared to HepG-2 spheres.Discussion and Conclusion: The anticancer activity is attributed to the bioactive compounds. The C. siphonella extract’s ability to induce apoptosis, disrupt mitochondrial membrane potential, and arrest the cell cycle highlights its potential as a novel anticancer agent. Additional research is required to investigate the underlying mechanism by which this extract functions as a highly effective anticancer agent. Keywords: Red Sea sponge, Callyspongia siphonella, breast cancer cell lines, hepatic cancer cell line, 2D and 3D cultures, cytotoxic activity

Keywords

• red sea sponge
• callyspongia siphonella
• breast cancer cell lines
• hepatic cancer cell line
• 2d and 3d cultures
• cytotoxic activity