Abstract: To study the antitumor activities of the constituents of Cephalotaxus fortunei distributed in Guizhou province. Methods ：The constituents were isolated by column chromatography and identified by physical and spectral analysis.Meanwhile，the anti-tumor activities of some compounds were evaluated by sulforhodamine B(SRB) and MTT assay. Results ：Eleven compounds were isolated and identified as apigenin (I)，β-sitosterol (II)，acetylcephalotaxine (III)，chrysoeriol (IV)，drupacine (V)，1-hentriacontanol (VI)，7，3′，4′-trihydroxyflavone (VII)，sugiol (VIII)，cephalotaxine (IX)，wllsonine (X)，and hainanolide (XI)，respectively.Biological screening Results demonstrated that some of the tested compounds exhibited the antitumor activities in vitro. Conclusion ：Compounds II，VI-VIII were isolated from this plant for the first time.Compound XI has a better inhibitory activity on cell line A549 and K562 .