Abstract

Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality worldwide, with limited treatment options in advanced stages. Traditional medicine has long provided alternative therapeutic avenues, particularly in Southeast Asia. YA-KAE-KASAI-LIN-KRA-BUE (KLB), a formulation found in the ancient Phaetsat Songkhro scripture of Thai traditional medicine, has been historically used to treat “Kasai,” a term referring to chronic diseases, including liver disorders. This study investigates the anti-HCC potential of KLB, assessing its antioxidant activity, cytotoxicity against HepG2 liver cancer cells, and biomolecular changes through advanced analytical methods, including gas chromatography-mass spectrometry and FTIR profiling. The KLB formulation demonstrated significant antioxidant properties, with Garcinia hanburyi and Senna siamea contributing the highest levels of phenolics and flavonoids, correlating with enhanced antioxidant activity. Cytotoxicity assays using Sulforhodamine B (SRB) revealed that KLB satu exhibited potent anti-proliferative effects on HepG2 cells at concentrations as low as 0.2–0.4 μg/ml, surpassing the efficacy of gemcitabine at similar dosages. Mechanistic studies showed that KLB induced S-phase cell cycle arrest and promoted apoptosis in HepG2 cells. FTIR analysis of ethanolic KLB satu extract highlighted significant biomolecular alterations, including increased lipid content and reduced nucleic acid/DNA intensity. Structural changes in proteins, particularly the shift of the β-sheet peak (1635 cm-1) and the emergence of an α-helix structure at 1654 cm-1, suggest potential alterations in protein conformation linked to apoptotic processes. These findings underscore the therapeutic potential of KLB as a natural agent for liver cancer treatment, providing scientific validation for its traditional use. The study also highlights the relevance of combining traditional knowledge with modern scientific techniques for the discovery of novel anti-cancer agents.

Key words: Anti-cancer, Gas Chromatography-Mass Spectrometry, Fourier Transform Infrared Spectrometer, Hepatocellular carcinoma, Thai Traditional Medicine