Identifying Antihepatocellular Carcinoma Compounds in Gansui Banxia Decoction Using Live Cell Adsorption

Gansui Banxia Decoction (GSBXD) is a traditional Chinese medicine formula for the treatment of hepatocellular carcinoma (HCC). Preliminary studies have identified the constituents and anticancer efficacy of GSBXD, but there is a gap in the screening of its lead compounds. Using live cell affinity combined with solid-phase extraction (LCA-SPE) and virtual screening and in vitro activity assays, we obtained 14 reliable potential lead compounds.

Coculturing H22 mouse HCC cells with GSBXD ethanol extract, isolating and purifying the bioactive fractions using LCA-SPE, and identifying the unknown bioactive components by ultraperformance liquid chromatography coupled with quadrupole time-of-flight full information tandem mass spectrometry (UPLC-QTOF-MS^E) with the UNIFI information processing platform were performed. Network pharmacology and molecular docking techniques predicted the potential mechanisms of these compounds against HCC. The enzyme-linked immunosorbent assay was used to examine the effects of core compounds on the expression of p53 and Bcl-2 in vitro.

Fourteen compounds screened from GSBXD using SPE-LCA-UPLC-QTOF-MS^E may be the main bioactive components. Network pharmacology predictions suggested that protein kinases regulate Bcl-2 and p53 expression to trigger the apoptosis in cancer cells. Molecular docking identified three core compounds, namely, lactiflorin, schaftoside, and violanthin, which showed high affinities for the relevant proteins. Experimental verification confirmed their superior anticancer activity in vitro. Their anti-HCC effects likely involved in the upregulation of p53 and downregulation of Bcl-2 expression at the cellular level.

We developed a stable and accurate SPE-LCA method, which successfully isolated and characterized 14 potentially active compounds from GSBXD. They may improve HCC by promoting p53 expression and reducing Bcl-2 expression. This work lays a foundation for discovering lead compounds and exploring potential mechanisms in traditional Chinese medicine formulas.