Two exopolysaccharides, NPS and APS, were isolated from L. plantarum WLPL09 and purified by ion-exchange chromatography. The structural analyses showed that molecular weight of NPS and APS were 72.60 kDa and 33.22 kDa, respectively. NPS was mainly composed of mannose and glucose, in molar ratio of 85.35:14.65, while APS was composed of mannose, glucose and galactose, in molar ratio of 89.69:8.65:1.66. In in vitro antitumor assays, APS displayed strong anti-proliferative effect against HepG2 hepatocellular carcinoma cells and HCT-8 colon adenocarcinoma cells in a dose-dependent manner. Morphological and flow cytometry analyses revealed that APS strongly induced apoptosis of HepG2 and HCT-8, especially for HCT-8. Furthermore, APS significantly up-regulated the mRNA level of apoptosis-related genes in cancer cells, and remarkably improved the activities of caspase-3, -8 and -9 in HepG2, caspase-3 and -8 in HCT-8. These results suggest APS might be explored as a potential, natural antitumor agent for functional food.