The current study was designed to investigate the toxicity of 3Z-3-[((1)H-pyrrol-2-yl)-methylidene]-1-(1-piperidinylmethyl)-1, 3-2H-indol-2-one (Z24), a novel synthetic indolin-2-ketone small molecule compound, using toxicogenomic techniques (complementary DNA [cDNA] microarray). Bioinformatic analysis suggested that the main functions of genes with altered expression were consistent with liver cell regeneration, apoptosis, metabolism of energy and fat, and the death receptor (DR)-mediated apoptosis-signaling pathway. Death receptor 4, Bcl-2, Bcl-xl, caspase 3, and cytochrome C, which are involved in the DR-mediated apoptosis-signaling pathway, were altered after Z24 treatment as determined by Western blotting analysis. When hepatocarcinoma cell line (HepG2 cells) treated with Z24 at 0.248 mmol/L for 24 hours, DNA fragmentation reached a maximum, and examination of cell morphology showed typical signs of apoptosis. These results indicate that Z24 can initiate apoptosis in hepatocytes, which in turn causes hepatotoxicity. A possible toxicological mechanism is that apoptosis was induced in hepatocytes by initiating the DR-mediated signal transduction pathway. Apoptosis of hepatocytes might lead to impairment of energy and lipid metabolism and provoke hepatocyte necrosis or inflammation, resulting in hepatotoxicity.