【作者】 李万锋；

【导师】 范志民；

【作者基本信息】 吉林大学 ， 外科学， 2009， 博士

【摘要】 乳腺癌是女性最常见的恶性肿瘤之一。在我国占全身恶性肿瘤的7%~10%,并呈逐年上升趋势,部分大城市报告乳腺癌占女性恶性肿瘤之首位。乳腺癌的发生是多因素、多步骤的生物学现象,单因素和局限在基因水平的研究难以全面揭示乳腺癌的发生机制。随着后基因组时代的来临,从细胞整体角度分析细胞内蛋白变化的蛋白质组学以其特有的思维方法和技术手段在解决生物学重大问题上开始显示出强大的威力,双向凝胶电泳和质谱技术的联合应用为乳腺癌蛋白质组学的分析提供了研究平台。通过比较疾病状态下与正常生理状态下或疾病不同发展阶段的蛋白质表达水平上的差异,以期发现和鉴定出与疾病特异相关的蛋白质,这是比较蛋白质组学研究的经典思路。本实验沿用这一思路,利用差异蛋白质组学方法详细研究了5例乳腺癌患者癌组织样品,与癌旁腺体组织进行比较,筛选出共同差异蛋白质钙结合蛋白S100A11,并利用Western blotting和免疫组化技术对其进行初步的验证,发现双向凝胶电泳、Western blotting和免疫组化结果得到相同的结论:S100A11在乳腺癌组织中的表达高于在癌旁乳腺组织中的表达。并探讨了该蛋白与乳腺癌临床参数之间的关系,发现S100A11与乳腺癌的浸润程度有关,据文献调研结果,S100A11的表达失调可能与乳腺癌的发生发展相关,为乳腺癌的发病学、防治研究提供了新的线索。更多还原

【Abstract】 Breast cancer is one of the most common malignant tumors. According to the statistics data, there are 1,200,000 women suffering from breast cancer, which takes up 18% in all of the female malignant disease. The five years survival rate of breast cancer is 50%～60%, and approximately 50% relapse and metastasis after treatment. So far, it still lack of a specific biomarker for early diagnosis and monitoring prognosis. Therefore, it is a key point to find a biomarker for breast cancer with high sensitivity and specificity. With the coming of post-genome period, proteomics with its special thinking method and technology which analyze the alteration of intracellular protein shows its great power to solve biological problems, and it attracts more and more researchers’attention. The classic methodology of proteomics is to find a special disease related protein by comparing the difference of protein expressing level between physiological condition and disease stages. This study researched breast cancer according to the method by two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), and we found that the expression of S100 calcium binding protein A11 (S100A11) up-regulated in breast cancer, which was validate by Western blotting and IHC (immunohistochemistry). In the result of IHC, S100A11 expressed mainly in breast cancer cell and adjacent normal breast epithelial cell and which was the result of expressing difference among them. We also found that S100A11 was associated with malignant cell infiltration. It provides a novel clue for studying pathogenesis and prevention of breast cancer.Objective: To find out the related protein of breast cancer and verify this different protein S100A11, and discuss the relationship between this protein and other clinical parameters.Methods: Choose 5 cases of malignant tissue in breast carcinoma and pairing adjacent normal tissue, and abstract total protein of malignant tissue and pairing normal tissue. ImageMaster 2D and Adobe Photoshop were used to analyze image, and MALDI-TOF-MS and MALDI-TOF-TOF-MS were used as protein identification. Searching Mascost database to find out different expressing protein and verify results generated from 2-DE and MS by Western blotting and IHC, and detect the protein expressing level in breast cancer and adjacent tissue by IHC .Result:1. S100A11 expressing level in breast malignant tissue is much higher than in adjacent tissue by Western blotting.2. Immunohistochemistry shows that: positive signal of protein S100A11 is brown granules, which located in plasma of carcinoma cell and adjacent cell without expressing in interstitium. In 55 cases of breast malignant tissue, the positive rate of S100A11 expressing is 73.73%; and in 26 cases of adjacent tissue, the positive rate of S100A11 expressing is 42.31%.There were significant difference of S100A11 expressing level between malignant tissue and normal tissue (χ2=7.005,P<0.05).3. Immunohistochemistry shows that: In 55 cases of breast malignant tissue, the expressing of S100A11 shows no relationship with patients’age, size of tumour, TNM clinical stage, ER, PR, HER-2, metastatic lymph node (P>0.05). The positive rate of S100A11 in early infiltrating carcinoma is 33.33 %; while in invasive carcinoma without specific type, it takes up 77.55 %, which showed no significant difference.Conclusion1. We got the same result by 2-DE, Western blotting and IHC, that is: S100A11 expressing level in breast malignant tissue is much higher than adjacent tissue by Western blotting, 2-DE and MS verify the conclusion.2. S100A11 express mainly in the plasma of breast cancer cell and adjacent normal breast epithelial cell, therefore the different content of S100A11 among the tissue result from difference expressing level of mammary gland cell .3. The expressing level of S100A11 in breast carcinoma cell is associated with the degree of infiltration. According to documents study, the disturbance of S100A11 expressing is probably related to occurrence of breast cancer and its development.更多还原