氟尿嘧啶诱导MCF-7乳腺癌细胞凋亡的蛋白质组研究  

Proteomic Analysis of 5-Fluorouracil Induced Apoptosis in MCF-7 Human Breast Carcinoma Cells

作　　者：蔡江霞[1] 陈思颖[1] 张蔚鹏[1] 孙金钥[1] 王陶陶[1] 吕军[1] 董亚琳[1] 

机构地区：[1]西安交通大学医学院第一附属医院药学部临床药学室,西安710061

出　　处：《中国药学杂志》2014年第4期303-308,共6页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(30973673)

摘　　要：目的研究氟尿嘧啶诱导人乳腺癌细胞MCF-7凋亡的分子机制。方法采用四甲基偶氮唑蓝法测定不同浓度氟尿嘧啶对MCF-7细胞的生长抑制作用;透射电镜观察氟尿嘧啶处理前后MCF-7内部的形态变化;流式细胞术测定氟尿嘧啶引起的MCF-7细胞周期分布变化及凋亡情况;氟尿嘧啶作用MCF-7细胞后收集细胞,利用蛋白质组学分析技术,对氟尿嘧啶诱导的MCF-7细胞凋亡前后的细胞蛋白进行二维凝胶电泳图谱分析,串联飞行时间质谱仪进行差异蛋白质分析。结果通过对凋亡前后的蛋白质组二维电泳图谱分析,筛选了14个差异倍数较大的蛋白点进行质谱分析,12个蛋白点通过质谱得到了鉴定,主要涉及能量代谢蛋白、细胞骨架蛋白、肿瘤侵袭转移蛋白及细胞信号转导蛋白等。结论本实验通过多种分析手段,鉴定了氟尿嘧啶诱导人乳腺癌细胞MCF-7凋亡前后发生变化的12种蛋白质,其中参与细胞信号转导的GNB2L1蛋白及能量代谢蛋白ECHS1可能在肿瘤发生发展中起着重要的作用,在氟尿嘧啶诱导乳腺癌细胞凋亡中扮演着重要角色,可作为新的抗肿瘤作用靶点。OBJECTIVE To investigate the mechanism of 5-fluorouracil （5-Fu） induced apoptosis in MCF-7 human breast carci- noma cells. METHODS The effect of different concentrations of 5-Fu on MCF-7 cells was detected by MTT assay. Transmission e- lectron microscope was used to observe morphological change of MCF-7 cells. Cell cycle distribution and apoptosis were determined by flow cytometry. The proteomic approach of combined 2-DE and MALDI TOF/TOF MS was used to identify differential protein expres- sion of MCF-7 induced by 5-Fu. RESULTS At 48 h after 5-Fu（98.2 μmol · L^-1 ）treatment, 14 large difference protein spots were screened to analysis by MALDI TOF/TOF MS. Among them 12 proteins were appraised, which involved in energy metabolism, cy- toskeleton, tumor invasion and metastasis, signal transduction. CONCLUSION We effectively found 12 changed proteins in the process of 5-Fu induced apoptosis in MCF-7 human breast carcinoma ceils by proteomic techniques. In these proteins, signal transduc- tion protein GNB2L1 and energy metabolism protein ECHS1 may play an important role in tumor development and may be novel diag- nostic marker for breast carcinoma.

关 键 词：氟尿嘧啶 乳腺癌 凋亡 蛋白质组学 

分 类 号：R965[医药卫生—药理学]