Cytotoxicity of Mineral Pitch against HEp-2 and Saos 2 cell lines: in vitro and in silico studies

Abstract

Mineral Pitch is a dark-colored, tar-like biomineral resin that naturally oozes from the cracks of mountain rocks. In traditional medicine, it is widely used for its tonic, antioxidant, anti-inflammatory, and regenerative properties. Recent studies have suggested that Mineral Pitch may also possess anticancer effects. This study aimed to evaluate the cytotoxic effects of Mineral Pitch on HEp-2 and Saos2 cancer cell lines as well as HEK293 and HUVEC normal cell lines, and to investigate its chemical profile and ligand–protein interactions through molecular docking analyses. Cell viability assessments were performed in a dose-dependent manner using the XTT assay. The major identified compounds (-)-epicatechin, baicalin, and resveratrol) were subjected to molecular docking analyses with the 4W6E and 6GGD protein targets. According to the XTT assay results, Mineral Pitch exhibited a dose-dependent cytotoxic effect on cancer cells, with the lowest IC₅₀ value (17.21 mg/mL) observed in the Saos2 cell line. Among normal cells, HEK293 was found to be more resistant, while HUVEC showed higher sensitivity. SI values indicated that Mineral Pitch possesses limited but significant selectivity toward tumour cells. HPLC results revealed that Mineral Pitch contains high levels of phenolic compounds, such as epicatechin, baicalin, and resveratrol. Molecular docking analyses demonstrated that epicatechin and baicalin exhibited high binding affinity and low Ki values with the 4W6E target protein. Mineral Pitch exhibits cytotoxic effects on cancer cells, indicating its potential as an anticancer agent.

Keywords

• HPLC-DAD analysis
• HEp-2 cancer cells
• Mineral pitch
• Molecular docking
• Saos2 cancer cells

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