Effectiveness of Methanol Solvent Extraction for Red Macroalgae Acanthophora spicifera Antitumoric Activity

Introduction: Red macroalgae Acanthophora spicifera is commonly found in most parts of Indonesia coastal areas. However, information regarding their potential pharmaceutical uses remains largely unexplored. This study evaluates the antiproliferative activity of Acanthophora spicifera collected from Indonesian extracted with different solvents of ethanol and methanol. Methods: Cytotoxicity and antiproliferative activity of A. spicifera crude ethanol extract (AS-EtOH) and A. spicifera crude methanol extract (AM-MetOH) were evaluated with Lactate dehidrogenase (LDH) and Trypan blue exclusion assay in Human cervical cancer (HeLa) cells for 72h. Concentration of extracts treated in HeLa cells were 6.25 to 200 μg/mL. Viability staining with fluorescence stain Hoechst33342/PI was conducted to investigate apoptotic activity. Potential apoptotic activity of treatments was confirmed by DNA fragmentation assay. Results: AS-MetOH demonstrated high cytotoxicity in HeLa cells with IC₅₀ of 127.3 ± 33.13 μg/mL. However, AS-EtOH IC₅₀ values could not be determined in this study. Antiproliferative activity was highly significant in AS-MetOH treated cells as cell density was suppressed to 20 x 10⁴ cells/mL compared to AS-EtOH (73 x 10⁴ cells/mL) and untreated cells (83 x 10⁴ cells/mL). Finally, apoptotic activity could be determined in AS-MetOH treated cells with increase PI fluorescence emitting cells and DNA fragmentation. On the other hand, apoptotic activity was not observable in AS-EtOH treated cells based on fluorescence viability staining and DNA fragmentation assay. Conclusion: Current results show methanol solvent as an effective solvent to promote potential pharmaceutical properties of macroalgae A. spicifera. Further advanced studies in a compound level from A. spicifera methanol fraction would be reasonable for development of macroalgae based anticancer agent.