腋窝淋巴结转移与无转移乳腺浸润性导管癌患者癌组织中差异表达蛋白的筛选与验证  

Screening and identification of differentially expressed proteins in patients with axillary lymph node metastasis and non-metastatic breast invasive ductal carcinoma

作　　者：杨亮 许文婷 赵倩 吴涛 迪力夏提·金汗 李丹 杜露 王波 朱丽萍 YANG Liang;XU Wenting;ZHAO Qian;WU Tao;Dilixiati·Jingsihan;LI Dan;DU Lu;WANG Bo;ZHU Liping(Cancer Hospital Affiliated to the Third Teaching Hospital of Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学第三临床医学院附属肿瘤医院

出　　处：《山东医药》2019年第29期5-8,共4页Shandong Medical Journal

基　　金：新疆维吾尔自治区自然科学基金资助项目(2016D01C353)

摘　　要：目的采用蛋白质组学技术分析乳腺浸润性导管癌组织的蛋白谱,寻找腋窝淋巴结转移与无转移患者的差异表达蛋白。方法选择行乳腺癌改良根治术的原发乳腺浸润性导管癌患者12例,其中腋窝淋巴结转移7例,腋窝淋巴结未转移5例。术中留取乳腺癌组织、癌旁组织和正常乳腺组织标本,提取组织样本中的蛋白,采用二维凝胶电泳法分析淋巴结转移和未转移乳腺癌组织的差异表达蛋白位点,蛋白表达量的差异倍数>2.0或<0.5且具有重复性时认为是差异表达蛋白位点(以癌旁组织和正常乳腺组织标本做对照,排除个体差异的影响);采用二维凝胶电泳结合基质辅助激光解吸电离飞行时间质谱对筛选出来的差异蛋白位点进行质谱鉴定。另取164例乳腺浸润性导管癌组织(其中腋窝淋巴结转移66例、无腋窝淋巴结转移98例),采用免疫组化SP法检测腋窝淋巴结转移与无转移患者癌组织中的差异表达蛋白。结果12例乳腺癌组织中,腋窝淋巴结转移和未转移乳腺癌组织共检出差异表达蛋白87个,其中差异表达最显著的6个蛋白分别是核基质结合区结合蛋白1(SATB1)、埃兹蛋白(Ezrin)、微管解聚蛋白(Stathmin)、E-钙黏附蛋白(E-cadherin)、Maspin蛋白、肿瘤转移抑制基因蛋白(KAI-1)。164例乳腺浸润性导管癌组织中,腋窝淋巴结转移乳腺癌组织Ezrin、KAI-1阳性表达率均高于未转移乳腺癌组织,E-cadherin蛋白阳性表达率低于未转移乳腺癌组织(P均<0.05)。结论乳腺浸润性导管癌腋窝淋巴结转移与无转移患者癌组织中Stathmi、KAI-1、E-cadherin蛋白表达存在差异,这些蛋白可能在淋巴结转移过程中发挥一定作用,可能作为评价乳腺癌淋巴转移的标志物。Objective The protein profiles of breast invasive ductal carcinoma patients were analyzed to find the differentially expressed proteins between axillary lymph node metastasis and non-metastasis by proteomic technology.Methods Twelve patients with breast invasive ductal carcinoma were selected.All patients underwent modified radical operation,including 7 cases of axillary lymph node metastasis and 5 cases of axillary lymph node non-metastasis.The samples of breast cancer and adjacent tissues were collected during the operation.Two-dimensional gel electrophoresis was used to analyze the differentially expressed protein sites in lymph node metastasis and non-metastasis breast cancer tissues.When the differential multiple of protein expression was greater than 2.0 or less than 0.5 and had reproducibility,it was considered as the differential expression protein locus(excluding the influence of individual differences by comparing the paracancerous tissues with the normal breast tissues).Two-dimensional gel electrophoresis combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry was used to identify the selected differential protein sites.In addition,164 cases of breast invasive ductal carcinoma(66 cases of breast cancer with lymph node metastasis,98 cases without axillary lymph node metastasis)were examined by immunohistochemical method to verify the differential expression of proteins in axillary lymph node metastasis and non-metastasis patients.Results Eighty-seven differentially expressed proteins were screened out in axillary lymph node metastatic breast cancer and non-metastatic breast cancer tissues.Six differentially expressed proteins were special AT rich sequence binding protein(SATB1),and Ezrin,Stathmin,E-cadherin,Maspin,and Kang-ai 1(KAI-1),respectively.In 164 cases of breast invasive ductal carcinoma,the positive expression rates of Stathmin and KAI-1 in the axillary lymph node metastatic breast cancer tissues was higher than that in non-metastasis breast cancer tissues,and

关 键 词：乳腺癌 淋巴结转移 蛋白质组学 二维凝胶电泳 埃兹蛋白 微管解聚蛋白 钙黏附蛋白 

分 类 号：R735.9[医药卫生—肿瘤]