Abstract
Marine green algae undergo substantial metabolic shifts during laboratory culture, yet the underlying biochemical mechanisms remain poorly understood. This study aimed to investigate the green seaweed Acrosiphonia orientalis’s multi-approach anti-proliferative and immunomodulatory activities by comparing extracts from laboratory-acclimatized (LabAO) and wild-harvested (WildAO) seaweed samples against human colorectal carcinoma (HCT-116) cell line. Higher accumulation of stress-response metabolites, including phytol, protocatechuic acid, neophytadiene, DHA, and EPA was observed in wild type extract (WildAO). In contrast, extract of lab-acclimatized biomass (LabAO) showed higher accumulation of storage metabolites like trehalose, maltose, and monoolein, as well as essential amino acids such as phenylalanine and isoleucine. Specifically, LabAO had more total sugars, phenolics, chlorophyll, and carotenoids, while WildAO had a higher concentration of flavonoids and proteins. Cancer-linked gene expression analysis revealed that both extracts triggered apoptosis as elevated PRKAA1 and IRS1 gene expression while suppressed SLC2A2 expression, leading to starved cells for energy. Further, GPX1 and SOD2 gene expression declined supporting oxidative stress. High expression of CASP3, TP53 and NDRG1 gene expression showed that cells pushed toward programmed death. Interestingly, expression pattern of NFKB1, MYC and BCL2 showed immunomodulatory activity. These findings highlight the potential of anti-proliferative and immunomodulatory activity, which may further explore for pharmaceuticals and nutraceuticals. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1038/s41598-026-39863-z.