Ovarian cancer is the most lethal gynaecologic malignancy, with a high mortality rate that is associated with the difficulty of treating the disease. This is due to the typical onset of symptoms when the disease is at a fairly advanced stage. Improved treatment strategies are required to improve longevity in patients. Recently polyphenols, secondary metabolites in plants, have aroused interest with respect to the treatment of cancer. The purpose of this study was to investigate the effect of two polyphenols, resveratrol and its derivative pterostilbene, on cell metabolism and proliferation in two ovarian cancer cell lines, OVCAR-5 and SKOV-3 that were grown in a three dimensional culture. The proteins VEGF, AKT, EGFR, HER2, cyclin D2 and PCNA were investigated to identify any possible changes to signalling pathways following exposure to the polyphenols. Both resveratrol and pterostilbene affected spheroid metabolism and proliferation in a dose- and cell line-dependant manner. Vascular endothelial growth factor and expression of the various proteins were affected following treatment. This study demonstrates that resveratrol and pterostilbene are capable of down regulating the cell metabolism and inhibiting proliferation in the ovarian cancer spheroids, potentially through several different pathways.