摘要
目的 :研究阻断泛素 -蛋白酶体通路对食管癌细胞生长及端粒酶活性的影响。方法 :将不同浓度的MG - 132加入食管癌细胞株Eca970 6 ,特异性阻断其泛素 -蛋白酶体通路 ,用四甲基偶氮唑蓝 (MTT)法测定细胞生长抑制效应 ,显微镜下观察细胞形态变化 ,流式细胞仪 (FCM)检测细胞周期及凋亡 ,DNA片段分析进一步证实凋亡的存在 ,并对端粒酶的活性进行检测。结果 :MG - 132对食管癌细胞株Eca970 6有显著的生长抑制作用 ,且呈现剂量与时间的依赖性 ,显微镜下可见明显的细胞病变 ,细胞变圆、变小、脱落 ,FCM显示MG - 132作用于食管癌细胞后 ,G1期比例增加 ,并有明显的亚二倍体凋亡峰 ,细胞DNA抽提电泳后发现凋亡特征性梯状条带 ,端粒酶的活性显著受抑制。结论 :MG - 132能显著抑制食管癌细胞株Eca970 6的生长 ,并抑制端粒酶的活性 ,表明阻断泛素
AIM: To investigate the effect on growth and activity of telomerase in esophageal carcinoma cells by inhibiting ubiquitin-proteasome pathway(UPP). METHODS: The esophageal carcinoma cell strain Eca9706 was treated with MG-132 to inhibit its UPP specially. The effect of growth suppression on cells was evaluated with MTT assay, morphologic changes of cells were observed under microscope, cell cycle and apoptosis were detected by flow cytometry (FCM). DNA fragment analysis was used to confirm the presence of apoptosis. The activity of telomerase was detected. RESULTS: MG-132 had obvious inhibitory effect on the growth of Eca9706 cells in a dose and time-dependent manner. Obvious pathologic change of cells were observed under microscope, cells became round, small and exfoliating. The FCM analysis showed that the ratio of esophageal carcinoma cells of G_1 phase increased and a obviously apoptotic sub-G_1 peak was found. Agarose electrophoresis showed marked ladder. The activity of telomerase was obviously inhibited. CONCLUSIONS: MG-132 significantly inhibits the growth and the activity of telomerase of Eca 9706 cells. These findings indicate that inhibiting UPP is a new strategy for the treatment of esophageal carcinoma. [
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第7期1208-1212,共5页
Chinese Journal of Pathophysiology
基金
本院重点科研资助课题
