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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 compare the mitochondrial protein fingerprints between cultured breast cancer cells and normal breast cells and screen the differential proteins to lay a foundation for the study of breast cancer subcellular protein markers. Methods The mitochondrial proteins of breast cancer cell lines MDA-MB-231, MCF-7 and normal breast cell line HBL-100 were extracted and analyzed by surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF- Statistical results, screening of differential proteins, and explore the impact of solvent and long-term cryogenic analysis of the results. Results Nearly 200 protein peaks were detected in the range of molecular weight from 2000 to 100 000. There were 45 and 36 protein peaks in HBL-100, MDA-MB-231 and MCF-7 cells, respectively (P < 0.05). The protein with relative molecular mass of 9 200, 13 800, 14 000 and 22 500 decreased in both cancer cells, and the protein with relative molecular mass of 10 000 and 11 600 in both cancer cells was increased . 4 ‰ of TritonX-114 and frozen storage for 3 to 6 months had no effect on the result analysis. Conclusion SELDI-TOF-MS can detect and analyze the mitochondrial differentially expressed protein in breast cancer rapidly and sensitively, which provides a new idea for the study of sub-cellular level breast cancer markers.