Assessing mechanism of action and effect of sphaerococcus coronopifolius extract in canines cell lines

The increasing focus on marine environments for drug research has led to more discoveries of compounds and drugs. This trend has been driven by green pharmacology, which emphasizes the use of algae and algae extracts. Specifically, the extract from Sphaerococcus coronopifolius has been found to have a positive effect in inhibiting the proliferation of certain canine cancer cell lines. This master's thesis project aimed to understand the mechanisms of action and effects of the extract from the alga Sphaerococcus coronopifolius on various canine cell lines, with the goal of understanding its potential as an anti-tumor therapy. The study employed GC/MS and HPLC/UV/MS to identify the main compounds present within the extract. Moreover, through this study we conducted toxicity assessment using the Alamar Blue assay on cancerous (C2 and NI-1 cell lines) and non-cancerous (Cf2Th and MDCK cell lines) canine cell lines, aiming to evaluate the extract's effects on both cancerous and normal cells and its selectivity towards cancerous cells. Additionally, quantitative real-time PCR (qPCR) was performed to investigate the mechanisms of action and effects of the algal extract, focusing on understanding which genes are regulated and how the algal treatment impacts both cancerous and control cells. The findings confirm the presence of bioactive compounds known for their anticancer properties, like diterpenes and triterpenes. In the other hand, the extract demonstrated a mild selective toxicity towards cancer cell lines, as evidenced by the Alamar Blue assay results. Moreover, the RT-qPCR results showed modulation of key genes involved in cell cycle regulation and cancer progression. Notably, the extract downregulated the expression of genes such as RAD51, CCNB2, PLK1, and SQLE in cancer cells, suggesting potential anticancer activity. The upregulation of CDKN1A, JUNB, FOS, and TP53INP1 further supports the extract's influence on cancer-related pathways, indicating its ability to affect gene expression in a manner that could be therapeutically beneficial. However, the extract's effects on normal cells, particularly MDCK cells, highlight the need for further research to optimize its selectivity and efficacy in cancer treatment. Overall, these results underscore the potential of S. coronopifolius extract as a source of bioactive compounds with applications in canine cancer therapy.