【作者】 汤礼宾；

【导师】 刘凤英；

【作者基本信息】 中南大学 ， 妇产科， 2008， 硕士

【摘要】 目的:观察JAK激酶抑制剂AG490对人宫颈癌细胞株Hela细胞增殖、凋亡及Bcl-2表达的影响。方法:（1）培养人宫颈癌细胞系Hela细胞并在光学显微镜下观察活细胞生长情况并摄片。（2）Hela细胞在含有10μmol/l、20μmol/l、40μmol/l、80μmol/l的AG490中培养24h、48h、72h,应用光学显微镜动态观察细胞形态学变化;（3）通过四甲基偶氮唑蓝（MTT）比色法观察AG490对细胞增殖活力的影响,计算细胞生长抑制率和IC₅₀。（4）采用流式细胞仪（FCM）定量检测终浓度为20μmol/l、40μmol/l、80μmol/l的AG490作用Hela细胞48h后不同浓度组细胞的凋亡率,并分析AG490对细胞周期分布的影响。（5）采用终浓度为20μmol/l、40μmol/l、80μmol/l的AG490处理Hela48h,之后应用免疫细胞化学染色技术观察Hela细胞的凋亡相关基因Bcl-2表达的变化。各实验对照组均加入同等浓度的DMSO,未加入其他干预因素。结果:（1）AG490以时间—剂量依赖性方式显著地抑制人宫颈癌细胞系Hela细胞体外的增殖活性（p＜0.05）。（2）经终浓度分别为10μmol/l、20μmol/l、40μmol/lAG490处理Hela细胞36h后,细胞被阻滞于G1期,S期细胞比例（SFP）明显降低（p＜0.05）。AG490作用24小时后,细胞bcl-2蛋白的表达降低,并出现典型的细胞凋亡的形态学表现,作用时间到48小时,Hela细胞凋亡达高峰。结论:1.JAK激酶抑制剂AG490能有效抑制人宫颈癌Hela细胞的体外增殖,其作用呈时间—量效关系。2.AG490下调Bcl-2蛋白表达,促进Hela细胞凋亡,凋亡率呈浓度依赖性。更多还原

【Abstract】 Objective: To investigate the effect and its mechanisms of AG490 on the proliferation, apoptosis and Bcl-2 expression of human cervical carcinoma cell lines Hela cells in vitro.Methods: Human cervical carcinoma cell lines Hela cells were cultured with different concentration of AG490 from 24 to 72 hours. The light microscopy was used to observe the changes morphologies of Hela cells.Methyl thiozolyl tetrazolium(MTT) was used to observe the growth suppressive rate of Hela. The flow cytometry (FCM) was used to investigate the phase of the proliferative cycles of Hela cells. The expression of Bcl-2 genes related to apoptosis were determinded by immunohistochemistry.Results: The proliferation of Hela cells were suppressed by AG490 in time-dose dependent fashion in vitro(p<0.05).Hela cells were blocked at Glphase,and the S-phase fraction(SPF) of the cells were decreased. When the concentration of AG490 was increased to 20umol/l for 24 hours,the expression of bcl-2 proteins decreased and the typical morphological changes of apoptosis appeared at 48 hours by AG490 treatment.更多还原