To study the chemical constituents of Piper wallichii (Miq.) Hand.-Mazz., and to explore the effects of extractive on proliferation and telomerase activity of human tongue squamous cell carcinoma Tca83 cells and its anti-cancer mechanisms. Sephadex LH-20 and several other column chromatography techniques were employed for separation of chemical constituents from Piper wallichii (Miq.) Hand.- Mazz., and for structure elucidation. Effects of different concentrations of extractive on Tca-83 cell proliferation were detected by MTT assay; meanwhile, changes in telomerase activity of Tca83 cells were determined by TRAP-ELISA. Data were statistically analyzed using SPSS 13.0. Five compounds were isolated and identified from ethyl acetate fraction of Piper wallichii (Miq.) Hand.-Mazz. Extractive could markedly inhibit Tca-83 cell proliferation in a dose and time dependent manner, and lower the telomerase activity. Extractive can markedly inhibit the proliferation and telomerase activity of Tca83 cells. Inhibition of telomerase activity may be one of the anti-tongue carcinoma mechanisms of Piper wallichii (Miq.) Hand.-Mazz.