摘要
目的研究雷公藤甲素对宫颈癌Hele细胞增殖的抑制作用。方法采用MTT方法检测雷公藤甲素对体外培养的宫颈癌Hele细胞增殖的影响,运用流式细胞术检测细胞周期,RT-PCR分析细胞周期蛋白B1(cyclin B1)mRNA的表达;Western-blot检测cyclin B1、P34^(cdc2)和磷酸化P34^(cdc2)(phosphor cdc2)蛋白的表达。结果雷公藤甲素浓度依赖性(0.5～4.0μg·mL^(-1))抑制体外培养的宫颈癌Hele细胞,干扰Hele细胞周期,细胞周期阻滞于S期和G_2/M期,S期和G_2/M期的百分率上升;同时G_0/G_1细胞百分率降低;RT-PCR结果显示,雷公藤甲素(0.5～4.0μg·mL^(-1))显著抑制cyclin B1 mRNA的表达;Western-blot结果显示,雷公藤甲素(0.5～4.0μg·mL^(-1))作用24h后cyclin B1蛋白表达开始不同程度降低,P34^(cdc2)变化较小,雷公藤甲素作用12h后磷酸化P34^(cdc2)蛋白水平明显改变。结论雷公藤甲素可以显著抑制宫颈癌Hele细胞体外增殖,其机制与细胞周期阻滞于S期和影响细胞周期因子cyclin B1的表达和改变P34^(cdc2)磷酸化有关。
Objective To study the effect of mptohde on proliferation of cervical cancer Hele cells in vitro and its related mechanisms. Method Inhibitory effect of cell proliferation was evaluated by MTF method; Cell cycle distributions were measured by flow cytometric assay; RT-PCR was used to analyze the expression of cyclin B 1 mRNA; Western blotting analysis was used to detect the protein expression of cyclin B1 ,P34cdc2 and phorylation P34cdc2. Results Triptolide （0.5-40 μg-mL-1） suppressed proliferation of cervical cancer Hele cells in a concentration-dependent manner. Flow cytometric analysis indicated that triptolide induced the cell cycle arrest at S and G2/M phase concentration-dependently at 24 h. Triptohde （0.5-4.0/.tg.mL-l） reduced the expression of cyclin B 1 mRNA. Western blotting analysis showed that treatment of Hele cells with triptohde remarkably decreased the expression of cyclin B1 at 24 h and enhancedthe level of phosphorylation P34cdc2 at 12 h. Conclusion Triptolide can significantly inhibit proliferation of the cervical cancer Hele cells and induce cell cycle arrest at the S and phase, and this induction is associated with the influence on expression of eyclin B 1 and phosphorylation P34cdc2 protein.
出处
《肿瘤药学》
CAS
2011年第1期33-36,共4页
Anti-Tumor Pharmacy
基金
湖南省科技计划项目资助
项目编号(2010SK3016)
