Objective To investigate the effect of phosphate-dayliositol-3-kinase/protein kinase B（PI3K/Akt） signaling pathway on Pien Tze Huang induced apoptosis of human ostesosarcom cell line U2OS. Methods U2OS cells were treated by different concentrations of Pien Tze Huang（0.4， 0.8， 1.2mg/mL）. The anti-proliferative effect of Pien Tze Huang on U2OS cells was measured by MTT. The apoptosis was detected by Hoeschst 33258 staining. The expression levels of the regulatory subunit of PI3K p85α（PI3K p85α）、 phospho-PI3K p85α（p-PI3K p85α）、 protein kinase B（Akt）、 phospho-Akt（p-Akt）、 Poly（ADP-ribose）polymerase（PARP）、 the cleavage of PARP（Cleaved PARP） were analyzed by Western blot. Results Pien Tze Huang suppressed the proliferation of U2OS cells in a time and dose-dependent manner. Topical morphological changes were observed by Hoechst 33258 staining. The expressions of p-PI3K p85α、 p-Akt were reduced and Cleaved PARP expression was increased while indicating a statistically significant difference as compared with the blank control group（P<0.05）. Conclusion Pien Tze Huang induces apoptosis of human osteosarcoma cell line U2OS through inhibiting PI3K p85αand Akt phosphorylation in PI3K/Akt signaling pathway and increasing the expression of Cleaved PARP.