熊胆粉对肝癌移植瘤裸鼠血管生成和细胞周期的影响

目的探讨熊胆粉对肝癌移植瘤裸鼠血管生成和细胞周期的影响。方法用人肝癌HepG2细胞建立裸鼠移植瘤模型,待皮下移植瘤形成后,将裸鼠随机分为熊胆粉组和对照组,每天分别给予熊胆粉和生理盐水灌胃,日1次,连续21 d。3周后剥取肿瘤。逆转录-聚合酶链反应(RT-PCR)检测p21、细胞周期蛋白依赖激酶2(CDK2)、细胞周期蛋白(CyclinE)、血管内皮生长因子(VEGF)、血管内皮生长因子受体(VEGFR2)mRNA的表达;免疫组化检测p21、CDK2、CyclinE、VEGF、VEGFR2蛋白表达,并通过CD31表达检测微血管密度(MVD)。结果 MVD结果显示熊胆粉显著抑制体内微血管的形成,与对照组比较差异有统计学意义(P<0.01);RT-PCR和免疫组化结果表明,熊胆粉显著促进p21的表达,抑制CDK2、CyclinE、VEGF、VEGFR2的表达,与对照组比较差异有统计学意义(P<0.01)。结论熊胆粉诱导细胞周期阻滞和抑制血管生成可能是其治疗肝癌的作用机制之一。

Impacts of Xiongdanfen on Angiogenesis and Cell Cycle in the Nude Mice with Hepatoma Xenograft

Objective To explore the impacts of xiongdanfen on angiogenesis and cell cycle in the nude mice with hepatoma xenograft. Methods The human hepatoma HepG2 cells were used to establish the model of xenograft in nude mice. When the subcutaneous xenograft was formed, the nude mice were random ized into an experimental group and a control group. Xiongdanfen and physical saline were used for gastric perfusion respectively every day,once a day,continuously for 21 days. In 3 weeks,the tumor was separated. RT - PCR was adopted to detect the expressions of p21, CDK2, CyclinE, VEGF and VEGFR2 mRNA. IHC method was used to detect the expressions of p21, CDK2, CyclinE, VEGF and VEGFR2 proteins. The mi crovessel density （MVD）was detected via CD31 expression. Results MVD result showed that xiongdanfen inhibited significantly the formation of capillaries in the body, presenting the significant difference as com pared with the control group（ P 〈 0.01 ）. RT- PCR and IHC results indicated that xiongdanfen promoted sig- nificantly p21 expression and inhibited the expressions of CDK2, CyclinE ,VEGF and VEGFR2 ,presenting the significant difference as compared with the control group （ P 〈 0.01 ）. Conclusion Xiongdanfen induces the blockage of cell cycle and inhibits angiogenesis ,which is possibly one of the effect mechanisms in the treatment of hepatoma.