乳腺癌细胞膜蛋白的SELDI-TOF-MS分析

Screening differential membrane proteins of cultured breast cancer cell lines

目的比较乳腺癌细胞株和正常乳腺上皮细胞膜蛋白指纹图谱,为乳腺癌亚细胞蛋白组学的研究奠定基础。方法试剂盒提取两株乳腺癌细胞株MDA-MB-231,MCF-7及一株乳腺正常上皮细胞HBL-100的膜蛋白,SEL-DI-TOF-MS分析蛋白指纹图谱,寻找差异蛋白。结果在分子量2kD-100kD范围内3株细胞均可检测到近100个蛋白点,MDA-MB-231和MCF-7与HBL-100相比,膜蛋白差异点分别为32个和34个(P<0.05),其中7.8kD,8.3kD和8.8kD的蛋白在两株癌细胞中表达均降低,9.6kD的蛋白在两株癌细胞中表达均增高。结论 SELDI-TOF-MS可以快速灵敏地检测到乳腺癌亚细胞蛋白水平的差异,为乳腺癌早期诊断提供依据。

Objective To screen differential membrane proteins by comparing protein fingerprinting between breast cancer and normal breast epithelial cell lines cultured in vitro,which may lay a foundation for sub-cellular protein biomarker of breast cancer.Methods Membrane proteins of two breast cancer cell lines MDA-MB-231,MCF-7 and one normal breast epithelium cell line HBL-100 were extracted,and the protein fingerprinting were measured by surface enhanced laser desorption/ionization time-of-flight mass spectrometry（SELDI-TOF-MS） for 50 times.Differential proteins were analyzed and screened by Bio-maker Wizard Software.Results About 100 protein peaks were observed on protein fingerprinting spectra ranged from 2 kD to 100 kD,among which 32 and 34 membrane protein peaks of them showed significant difference between MDA-MB-231 and MCF-7 when compared with HBL-100（P〈0.05）.Protein peaks at 7.8 kD,8.3 kD and 8.8 kD were down-regulated in both breast cancer cell lines,while protein peaks at 9.6 kD were up-regulated.Conclusion SELDI-TOF-MS can detect membrane proteins of breast cancer cell lines accurately and sensitively,which applies a new method for subcellular protein biomarkers of breast cancer.