| Summary: | <i>Indigofera zollingeriana</i> Miq (<i>I.</i> <i>zollingeriana</i>) is a widely grown tree in Vietnam. It is used to cure various illnesses. The purpose of this study was to investigate the chemical constituents of an <i>I. zollingeriana</i> extract and test its anticancer activity on hepatocellular cells (Huh7 and HepG2). The experimental results of the analysis of the bioactive compounds revealed that β-sitosterol (β-S) and β-sitosterol-glucoside (β-SG) were the main ingredients of the <i>I.</i> <i>zollingeriana</i> extract. Regarding anticancer activity, the β-S and β-SG of <i>I. zollingeriana</i> were found to exhibit cytotoxic effects against HepG2 and Huh7 cells, but not against normal human primary fibroblasts. The β-S was able to inhibit the proliferation of HepG2 and Huh7 cells in a dose-dependent manner with half-maximal inhibitory concentration (IC<sub>50</sub>) values of 6.85 ± 0.61 µg/mL and 8.71 ± 0.21 µg/mL, respectively (<i>p</i> < 0.01), whereas the β-SG IC<sub>50</sub> values were 4.64 ± 0.48 µg/mL for HepG2 and 5.25 ± 0.14 µg/mL for Huh7 cells (<i>p</i> < 0.01). Remarkably, our study also indicated that β-S and β-SG exhibited cytotoxic activities via inducing apoptosis and activating caspase-3 and -9 in these cells. These findings demonstrated that β-S and β-SG from <i>I.</i> <i>zollingeriana</i> could potentially be developed into promising therapeutic agents to treat liver cancer.
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