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In this study, crude extracts of the marine eelgrass Zostera japonica and their solvent-partitioned fractions were evaluated for their inhibitory effect against AGS, HT-1080 and MCF-7 human cancer cells using MTT assay. Each of the crude extracts (acetone/methylene, chloride, and methanol) of Z. japonica showed a significant inhibitory effect on the growth of human cancer cells. The combined crude extracts were partitioned between CH2Cl2 and water. The organic layer was further partitioned between 85% aq. MeOH and n-hexane, and the aqueous layer was then fractionated into n-BuOH and H2O, successively. Growth inhibition effects of solvent-partitioned fractions from Z. japonica on human cancer cells increased in a dose-dependent manner. Among these tested samples, the 85% aq. MeOH fraction revealed good inhibitory effects on the growth of AGS and HT-1080 human cancer cells, while the n-hexane fraction exhibited good inhibitory effects on the growth of AGS and MCF-7 human cancer cells. In addition, 85% aq. MeOH and n-hexane fractions enhanced mRNA expression of p53 gene. These results suggest that there is further scope for the isolation of active compounds from Z. japonica, which should show much stronger anticancer activity.