体外培养乳腺癌细胞线粒体差异蛋白的筛选  

Screening of Differential Mitochondrial Protein in Breast Cancer Cells Cultured In Vitro

作　　者：韵雪雪[1] 杨永长[2] 姜伟[2] 肖代雯[2] 闫慧[1] 黄文芳[2] 罗春丽[1] 

机构地区：[1]重庆医科大学医学检验系临床检验诊断学教育部重点实验室,重庆400016 [2]四川省医学科学院四川省人民医院,成都610072

出　　处：《中国生物制品学杂志》2011年第5期571-574,共4页Chinese Journal of Biologicals

基　　金：国家"863"计划资助项目(2006AA02090408)

摘　　要：目的比较体外培养的乳腺癌细胞与正常乳腺细胞线粒体蛋白指纹图谱,筛选差异蛋白,为乳腺癌亚细胞蛋白标志物的研究奠定基础。方法提取乳腺癌细胞株MDA-MB-231、MCF-7及正常乳腺细胞株HBL-100线粒体蛋白,表面增强激光解析蛋白飞行时间质谱(SELDI-TOF-MS)检测蛋白表达,Biomarker Wizard Software分析软件统计结果,筛选差异蛋白,并探讨溶剂及长期冻存对结果分析的影响。结果在相对分子质量2 000～100 000范围内检测到近200个蛋白峰,HBL-100与MDA-MB-231和MCF-7细胞相比,差异蛋白峰分别为45个和36个(P<0.05),其中相对分子质量9 200、13 800、14 000和22 500的蛋白在2株癌细胞中表达均下降,相对分子质量10 000和11 600的蛋白在2株癌细胞中表达均升高。4‰的TritonX-114及冻存3～6个月对结果分析无影响。结论 SELDI-TOF-MS能够快速、灵敏地检测分析乳腺癌线粒体差异表达蛋白,为亚细胞水平乳腺癌标志物的研究提供了新的思路。Objective To screen differential mitochondrial protein by comparing the protein fingerprint of breast cancer cells cultured in vitro with that of normal breast cells,and lay a foundation of study on subcellular protein biomarker of breast cancer.Methods Mitochondrial proteins were extracted from breast cancer cell MDA-MB-231 and MCF-7 strains and normal breast cell HBL-100 strain,and analyzed for fingerprints by surface enhanced laser desorption ionization time-of-flight mass spectrometry（SELDI-TOF-MS）.The results were analyzed by Biomarker Wizard Software,based on which differential mitochondrial proteins were screened,and the effect of dissolvent and long-term storage in frozen on the result analysis was evaluated.Results Nearly 200 protein peaks were observed in a relative molecular mass range of 2 000 to 100 000.Compared with that of HBL-100 cells,45 and 36 differential protein peaks were observed in MDA-MB-231 and MCF-7 cells（P 0.05） respectively.The expression levels of proteins with relative molecular masses of 9 200,13 800,14 000 and 22 500 in the two breast cancer cell strains decreased,while those with relative molecular masses of 10 000 and 11 600 increased.Triton X-114 at a concentration of 4‰ and storage in frozen for 3 ～ 6 months showed no effect on result analysis.Conclusion SELDI-TOF-MS may be used for rapid and sensitive detection of differential mitochondrial protein in breast cancer cells,which provided a novel route for study on subcellular protein biomarker of breast cancer.

关 键 词：乳腺肿瘤 线粒体蛋白质类 表面增强激光解析蛋白飞行时间质谱 蛋白指纹图谱 差异蛋白 

分 类 号：R737.9[医药卫生—肿瘤]