目的 探讨重楼提取物（PPEE）抗骨肉瘤作用及其机制。方法 不同质量浓度的PPEE处理骨肉瘤细胞，采用MTT法、Hoechst 33342染色、流式细胞术、Matrigel培养、免疫印迹法分别检测细胞增殖抑制率、细胞凋亡、细胞周期、血管生成拟态（VM）形成及相关蛋白表达情况。裸鼠移植瘤模型观察PPEE的体内抑瘤效果。结果 PPEE对骨肉瘤细胞半数抑制浓度（IC₅₀）为10~60 μg/mL，对成骨细胞增殖影响较小。PPEE可浓度依赖性地将骨肉瘤143B细胞阻滞在G₂/M期，上调细胞周期相关蛋白p-CDK1、p-Cdc25C、p-Chk2表达，下调cyclin B1表达；促进细胞凋亡，上调cleaved Caspase-3、8、9和PARP表达，上调Bax/Bcl-2；抑制细胞体外VM形成，下调FAK、Mig-7、MMP-2和MMP-9表达。体内实验显示PPEE能明显抑制骨肉瘤生长和体内VM形成，延长荷瘤裸鼠生存期。结论 PPEE在体内外均具有良好的抗骨肉瘤活性，其作用机制可能与诱导骨肉瘤细胞凋亡、阻滞细胞周期及破坏骨肉瘤VM形成有关。

Objective To evaluate the potential anti-osteosarcoma effects of Paris polyphylla ethanol extract (PPEE) and investigate its underlying mechanisms.Methods The anti-proliferation activity of PPEE was tested on 143B, MG-63, U-2 OS, and hFOB1.19 cells using MTT assay. The pro-apoptotic and cell cycle arrest effects of PPEE were confirmed by Hoechst 33342 staining and flow cytometry. The anti-vasculogenic mimicry effects of PPEE were investigated by Matrigel culture assays. The related proteins expression was evaluated by Western blottig.Results PPEE evidently suppressed cell proliferation of 143B, MG-63, and U-2 OS cells with IC₅₀ values of 10-60 μg/mL, but showed little cytotoxicity against normal osteoblastic cell. PPEE induced G₂/M phase arrest associated with elevated phosphorylation of CDK1, Cdc25C, Chk2 and down-regulation of cyclin B1 expression. It also promoted apoptosis in 143B cells by up-regulating the expression of cleaved Caspase-3, Caspase-8, and Caspase-9, and Bax/Bcl-2 ratio. Additionally, PPEE inhibited 143B cells vasculogenic mimicry formation at non-cytotoxic concentrations through decreasing the expression of FAK, Mig-7, MMP-2, and MMP-9. Finally, four weeks' daily oral administration of PPEE exhibited potent antitumor and anti-vasculogenic mimicry activity in 143B xenograft model with low toxicity.Conclusion These findings demonstrated that PPEE possesses anti-osteosarcoma and anti-vasculogenic mimicry activity in vitro and in vivo, and its underlying mechanisms may be related to inducing apoptosis, blocking cell cycle, and destroying vasculogenic mimicry formation of osteosarcoma cells. Therefore, PPEE is a potential candidate for osteosarcoma treatment.

国家自然科学基金资助项目（81673017）；江苏省自然科学基金资助项目（BK2012775）