Apoptotic effects of crude muscle extract from marine pufferfish Chelonodon patoca against human cancer cell lines

Abstract Background The global establishment of cancer registries has prompted a hunt for innovative medications that destroy cancer but not healthy cells. Researchers are currently searching marine environments for new anticancer drugs. Modern chemotherapy uses numerous compounds of aquatic origin. These substances exhibit cytotoxic characteristics through various methods, including DNA damage, apoptosis induction, and growth suppression of cancerous cells. The discovery and development of novel anticancer agents from abundant marine fish is generating increasing interest. The objective of this present study is to extract and assess the anti-proliferative effect of a crude muscle extract from marine pufferfish on human cancer cell lines. Methods The biochemical constituents, protein profile, and anticancer activities of the prepared crude muscle extract were analysed using standard methods on human cancer cell lines (HT-29, MDA-MB-231, A-549, and AGS). Results The biochemical contents, such as protein (7.19 ± 0.20 mg/mL), lipid (1.56 ± 0.14 mg/mL), and carbohydrate (1.19 ± 0.09 mg/mL), were quantitatively analysed. Native PAGE and SDS-PAGE qualitatively analysed the protein profiles, revealing distinct protein bands with molecular weights ranging from 220 to 14 kDa. The crude muscle extract was screened for its cytotoxicity (vero) and cell viability (HT-29, MDA-MB-231, A-549, and AGS) against human cancer cell lines by the MTT assay method. The nuclear morphological changes of the apoptotic cells were stained using propidium iodide, and the morphological changes associated with apoptosis were assessed using acridine orange/ethidium bromide (AO/EB) fluorescence staining. The intensity of the mitochondrial membrane potential of the treated cells was measured using the Rh-123 stain, and the results of the DNA fragmentation assay showed that the crude muscle extract-treated cells showed DNA damage, which is indicative of apoptosis. Conclusions This preliminary study supports that the crude muscle extract from milk spotted marine pufferfish, C. patoca has strong anticancer properties. This implies potential for the development of more effective anticancer drugs in future. Graphical abstract