Mangiferolic acid from Thai Tetragonula laeviceps propolis triggers inflammation in Hep-G2 liver cancer cells via NF-κB and COX2 upregulation and G1 arrest

Liver cancer ranks among the leading causes of cancer mortality worldwide, with treatments limited by high recurrence and severe side effects. Here, the anticancer potential of mangiferolic acid (MF) from Tetragonula laeviceps propolis against liver cancer was evaluated. This in vitro study used Hep-G2 liver cancer cells. Cytotoxicity was investigated by the MTT assay. Morphological changes were observed under a compound microscope. Apoptosis, necrosis, and cell cycle distribution were analysed using annexin V/propidium iodide (PI) flow cytometry. Expression of CASP7, COX2, and NF-κB was quantified using RT-qPCR. MF demonstrated strong cytotoxicity against Hep-G2 cells, with IC₅₀ values of 8.35-18.41 µg/mL (18.28-40.31 µM). Morphological analysis showed dose- and time-dependent cell shrinkage and density reduction. Annexin V flow cytometry revealed increased late apoptotic and necrotic populations after 72 h at 30 µg/mL (65.69 µM). MF induced G1-phase arrest, with 70.24% of cells halted in G1, reducing S-phase progression. Gene expression analysis showed upregulation of CASP7, COX2, and NF-κB after 72 h, suggesting apoptosis induction and inflammatory necrosis. MF inhibited Hep-G2 cell proliferation, induced apoptosis, and caused cell cycle arrest, highlighting its potential for cancer inhibition in vitro. However, further research is necessary to clarify its role and evaluate its applicability.