采用表面增强激光解吸离子化飞行时间质谱技术筛选乳腺癌新辅助化疗敏感蛋白谱

目的应用表面增强激光解吸离子化飞行时间质谱（SELDI-TOF MS）技术检测乳腺癌新辅助化疗前后癌组织内蛋白质谱的变化,筛选乳腺癌新辅助化疗敏感性相关蛋白质。方法 60份样本来自2009年3月至2010年8月山东省千佛山医院乳腺外科收治的30例乳腺癌患者CEF新辅助化疗前后的乳腺癌组织。化疗前,行粗针穿刺获得乳腺癌组织;化疗后,在乳腺癌根治术中取样。应用SELDI-TOF MS技术检测乳腺癌组织的蛋白质谱,所得数据采用Biomarker wizard 3.0.2软件分析,得到差异蛋白谱,再采用SPSS17.0软件对数据进行Shapiro-Wilk正态性检验和配对设计t检验。根据差异蛋白质的质荷比,在SWISS-PROT数据库中检索与差异蛋白质相匹配的存在于乳腺组织或乳腺癌组织中的蛋白质。结果通过比较患者新辅助化疗前后的蛋白峰值,筛选出8个蛋白峰有差异的蛋白质,其质荷比（m/z）分别为5825.9、8949.5、11053.5、11652.0、17898.9、22250.6、26055.5、38546.6,化疗后这8个蛋白峰均高于化疗前（11.0±5.0比8.2±4.9、10.7±4.5比6.3±3.4、22.3±9.9比15.3±9.9、28.0±10.2比21.4±9.3、9.5±4.6比5.4±3.4、1.1±1.0比0.6±0.5、2.0±0.7比1.5±0.7及2.7±2.2比1.5±1.2,P值均〈0.05）。结论应用SELDI-TOF MS技术可以筛选出与乳腺癌新辅助化疗敏感性相关的蛋白质谱。

Screening the sensitive protein spectrum in breast cancer tissue before and after neoadjuvant chemotherapy by SELDI-TOF MS technology

Objective To detect protein spectrum of breast cancer tissue before and after neoadjuvant chemotherapy （NACT） using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry （SELDI-TOF MS）, in order to find the sensitive proteins to neoadjuvant chemotherapy. Methods We collected 60 samples of breast cancer tissues before and after NACT in 30 patients treated in Qianfoshan Hospital from March 2009 to August 2010. The breast cancer tissue sample was obtained by core needle biopsy before NACT and during radical mastectomy after NACT. SELDI-TOF MS was applied to detect the protein spectrum in breast cancer tissue and Biomarker Wizard 3.0.2 version was used for screening the differential protein spectrum. The data were analyzed by means of Shapiro-Wilk test and matching t test in SPSS17.0 software. According to the m/z of differential proteins, we retrieved in SWISS-PROT database and found the proteins in breast tissue or breast cancer tissue consistent with the differential proteins. Results In this study, 8 different proteins were screened in NACT group： m/z 5825.9, 8949.5, 11 053.5, 11 652.0, 17 898.9, 22 250.6, 26 055.5, 38 546.6. They were all expressed higher after NACT （11.0±5.0 vs 8.2±4.9, 10.7±4.5 vs 6.3±3.4, 22.3±9.9 vs 15.3±9.9, 28.0±10.2 vs 21.4±9.3, 9.5±4.6 vs 5.4±3.4, 1.1±1.0 vs 0.6±0.5, 2.0±0.7 vs 1.5±0.7 and 2.7±2.2 vs 1.5±1.2, P〈0.05）. Conclusion Protein spectrum which is sensitive to neoadjuvant chemotherapy in breast cancer can be screened by SELDI-TOF MS.