腋淋巴结阳性乳腺浸润性导管癌差异蛋白的质谱分析

A mass spectrometric analysis of differential proteins in breast infiltrative ductal carcinoma patients with axillary lymph node metastases

目的筛选可能用于判断乳腺浸润性导管癌发生腋淋巴结转移的血清蛋白标志物。方法收集2015年3月—2016年5月天津市宝坻区人民医院收治的乳腺浸润性导管癌患者血清样本24份,按照是否发生腋淋巴结转移分为两组,每组各12份。将每组的12份血清等量混合,利用双向凝胶电泳(2D-PAGE)和基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)技术鉴定差异蛋白质,对2D-PAGE图谱与MALDI-TOF-MS质谱进行分析。结果 2D-PAGE图谱分析筛选出差异蛋白点11个;经MALDI-TOF-MS质谱分析鉴定出高可信度的蛋白5种,表达上调的为T淋巴瘤侵袭转移诱导因子1(Tiam1)、组织蛋白酶K(Cat K)、血管内皮生长因子B(VEGF-B)、肌动蛋白相关蛋白复合物3(ARPC3),表达下调的为抑癌蛋白Necdin(覆盖率分别为Tiam1蛋白29%,Cat K蛋白12%,VEGF-B蛋白34%,ARPC3蛋白26%,Necdin蛋白18%)。结论乳腺浸润性导管癌发生腋淋巴结转移后其蛋白质表达谱也发生了变化,所鉴定出的差异蛋白有可能成为判断其是否发生腋淋巴结转移的血清蛋白标志物。

Objective To screen serum protein markers that may be used in diagnosis of axillary lymph node metastasis in patients with breast infiltrative ductal carcinoma. Methods From March 2015 to May 2016, 24 serum samples of patients with breast infiltrative ductal carcinoma from Baodi District People＇s Hospital of Tianjin were collected. All the samples were divided into two groups according to whether there was an axillary lymph node metastasis or not, and each group contained 12 samples. Equal amount of 12 samples in each group were mixed, respectively. The differential proteins were identified by using two-dimensional polyacrylamide gel electrophoresis（2D-PAGE） and matrix-assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF-MS） techniques, and the atlas of 2D-PAGE and MALDI-TOF-MS mass spectrum were analyzed. Results Eleven differential proteins＇ spots were screened out by 2D-PAGE analysis; by MALDI-TOF-MS mass spectrum analysis, 5 high-confidence proteins were identified, including the expressions of up-regulated T-lymphoma invasion and metastasis-inducing factor 1（Tiam1）, cathepsin K（Cat K）, vascular endothelial growth factor B（VEGF-B） and actinrelated protein complex 3（ARPC3）, and the down-regulated expression of tumor suppressor protein Necdin（Coverage： Tiam1 protein was 29%, Cat K protein was 12%, VEGF-B protein was 34%, ARPC3 protein was 26%, Necdin protein was 18% respectively）. Conclusion After axillary lymph node metastasis occurs in patients with breast infiltrative ductal carcinoma, its protein expression profile has also changed. The differential proteins identified may become the serum protein markers to determine whether there is an axillary lymph node metastasis or not.