From the methanolic extract of fresh fruits of Solanum reflexum Schrank, a major compound was isolated and identified as; aculeatiside-A 1. The structure of the isolated compound was elucidated on the basis of 1H and 13C and DEPT-13C-NMR; in addition to comparison with reported data. The cytotoxic effect of aculeatiside-A on hepatocellular carcinoma cells and its mechanism was investigated. It was accomplished that aculeatiside-A inhibit the growth of HepG-2 cells significantly by induction of apoptosis. Marked morphological changes of apoptosis were observed clearly. The results for the first time demonstrate that aculeatiside-A has significant anti-proliferation effect by induction of apoptosis on hepatocellular carcinoma cells in-vitro, down regulation of Survivin and Bcl-2 expressions as well as up regulation of Bax and P53 expression may be one of the important apoptotic inducing mechanisms.