紫杉醇诱导的乳腺癌耐药细胞的蛋白质组学研究  

Proteomic Analysis of Proteins Associated with the Paclitaxel-Induced Drug-Resistant Human Breast Cancer Cells

作　　者：陈思颖[1] 蔡江霞[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10期825-832,共8页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(30973673)

摘　　要：目的应用蛋白组学分析方法探讨紫杉醇诱导的人乳腺癌细胞的多药耐药机制。方法采用低浓度加量持续诱导法建立紫杉醇耐药的人乳腺癌细胞株(MCF-7/Tax);利用MTT法、流式细胞术研究细胞的生物学特性;应用二维凝胶电泳(2-DE)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析亲本细胞(MCF-7/S)耐药前后的差异蛋白;然后选取5种差异较大且具有不同功能的蛋白,利用Western-blot和Real-time PCR实验对其进行验证。结果成功建立紫杉醇耐药的人乳腺癌细胞株MCF-7/Tax,耐药倍数为115倍;质谱成功鉴定出MCF-7/Tax和MCF-7/S之间的17种差异蛋白,其中11种蛋白在MCF-7/Tax中表达上调,6种表达下调。涉及肿瘤发生和发展,能量代谢和细胞骨架功能的4种蛋白hnRNP C1/C2,SET,AAT及Transgelin-2均在MCF-7/Tax中表达上调,而与肿瘤侵袭转移相关的NDKA蛋白在MCF-7/Tax中表达下调,它们mRNA水平的验证结果与蛋白水平分析一致。结论鉴定出的5种差异蛋白,尤其是Transgelin-2为探讨乳腺癌的紫杉醇耐药机制提供新的依据,有望成为临床上治疗乳腺癌新的分子靶标。OBJECTIVE To investigate multidrug resistance（ MDR） in the paclitaxel-induced drug-resistant breast cancer MCF-7 cells（ MCF-7 / Tax） using proteomic analysis. METHODS MCF-7 / Tax cell line was established by escalating the concentrations of paclitaxel to drug-sensitive MCF-7 cells（ MCF-7 / S）. The biological characteristics of MCF-7 / Tax cells were analyzed using MTT test and flow cytometry. The global protein profiles of MCF-7 / Tax and MCF-7 / S were compared using two-dimensional gel electrophoresis（ 2-DE） and matrix-assisted laser desorption/ionization time of flight mass spectrometry（ MALDI-TOF-MS）. We confirmed the protein and mRNA levels of five differential patterns of expression by Western blot and Real-time PCR,respectively. RESULTS The resistance factor of MCF-7 / Tax was 115. Significant differential expressions of 17 proteins between MCF-7 / Tax and MCF-7 / S were identified with 11 proteins upregulated and six proteins downregulated in MCF-7 / Tax cells. With western blot and real-time PCR,we confirmed that heterogeneous nuclear ribonucleoprotein（ hnRNP C1 / C2）,SET nuclear oncogene（ SET）,aspartate aminotransferase（ AAT）, transgelin-2 were upregulated,and nucleoside-diphosphate kinase A（ NDKA） was downregulated in MCF-7 / Tax cells. CONCLUSION The identification of differential proteins,particularly transgelin-2 provides new insights into the mechanism of MDR to paclitaxel and novel biological targets for breast cancer treatment.

关 键 词：乳腺癌 紫杉醇 多药耐药 蛋白质组学 

分 类 号：R965[医药卫生—药理学]