乳腺癌為全世界女性發生率最高的癌症，每年有一百一十五萬新增病例，佔全世界每年癌症人數一千一百萬的百分之十。近年來，台灣每年也有將近六千名新增的乳腺癌病例，且發病年齡有年輕化之趨勢，好發年齡高峰期在四十五至五十五歲之間，與西方歐美國家相較年輕十歲。由於乳腺癌須依賴乳房超音波、乳房磁振掃描、傳統的病理組織學，作為篩檢、診斷及治療的預後相關因子外，尚需輔以生物腫瘤標記作為參考。基於以上所述特性，針對台灣婦女乳腺癌的研究，成為一個重要的課題。
    近幾年的文獻提到乳腺癌表現因子除了雌激素接受體(ER)、黃體激素接受體(PR)及腫瘤蛋白Her-2/neu和預後有相關以外，乳腺癌發生機轉也與生物發育中佔極重要角色的Notch signal pathway有關，其中Notch family接受體Notch1和與其結合之受質Jagged1、Delta之表現和腫瘤蛋白HER-2/neu之表現有關聯，因此我們想更進一步分析Notch1、Jagged1、Delta三者之相對表現差異。本研究利用組織微陣列工具，針對兩類型乳腺癌來建立組織晶片。而此兩類型區分，以病理報告中的乳腺癌表現因子ER/PR及腫瘤蛋白HER-2/neu陽性或陰性來區分，分為ER/PR陰性和HER-2/neu陽性，以及ER/PR陽性而HER-2/neu陰性兩大類，再以取出癌症組織建立組織晶片。利用所建立乳腺癌組織晶片，以Notch1、Jagged1、Delta之抗體進行免疫組織化學染色，所得之結果利用卡方檢定及Spearman等級相關係數檢定，統計分析。
    其結果呈現在59案例中，HER-2/neu的表現與組織惡性度有正相關性。而Notch1的表現量和組織惡性度呈現負相關性。Jagged1在乳腺癌組織的表現和Notch1相似。但Delta表現量則與Jagged1的表現呈現負相關。最後，Jagged1的表現與組織惡性度呈現負相關性。但是在HER-2/neu陰性條件下，Jagged1的表現與組織惡性度呈現正相關性。
關鍵字：雌激素接受體、黃體激素接受體、腫瘤蛋白HER-2/neu、卡方檢定、組織微陣列、組織惡性度

Breast cancer has the highest incidence in all the malignancy in the world. Around 1.15 million new cases of breast cancer are found every year, which are accounted for ten percent of the 11 million people newly-diagnosed with cancer. In recent years, there are near six thousand new cases of breast cancer occurring in Taiwan, and their age have a younger tendency. The peak age of breast cancer in Taiwan is around 45-55 years old, which is 10 years younger than that in the western countries. For screening, diagnosis and treatment of breast cancer, breast ultrasound, magnetic resonance imaging, and traditional histopathology are still the gold standard. Tumor markers such as estrogen receptor, progesterone receptor and oncoprotein Her-2/neu are also known as a guidance for prognosis. Therefore, the specific study of breast cancer in Taiwan women becomes an important issue.
   Recent studies suggest that breast cancer biomarkers are estrogen receptor (ER), progesterone receptor (PR) and oncogene protein Her-2/neu, and they are related to prognosis. Notch signal pathway, which plays important role in biological development, is also related to the pathogenetic mechanism of breast cancer. One of Notch family receptors, Notch1, and its ligands Jagged1 and Delta, the expression of are associated with tumor-related protein Her-2/neu expression. Therefore, we would like to further analyze the differential expression between Notch1, Jagged1 and Delta in different sets of breast tumors. In this study, we use tissue microarray as a tool to establish tissue microarrays for two different types of breast cancer. In this study, breast cancer is classified into two groups based on pathologic reports of ER, PR and Her-2/neu expression, positive and/or negative. The two groups are ER/PR negative and Her-2/neu positive, and ER/PR positive and Her-2/neu negative. Then we established tissue microarrays, which were taken from breast cancer tissue cores. Notch1, Jagged1, and Delta protein expression were evaluated by immunohistochemistry in tissue microarrays of breast caner. The association between the expression patterns of these proteins and biomarkers of breast cancer were analyzed by χ2 test and Spearman’s rank correction test.
   In the total 59 cases, expression of HER-2/neu and the tumor grading of invasive breast cancer shows positive correlation. The expression of Notch1 and tumor grade of invasive breast cancer presents negative correlation. The expression of Jagged1 in invasive breast cancer is similar to that of Notch1. Delta expression strength reveals negative correlation to the expression of Jagged1. Finally, expression of Jagged1 and tumor grade of invasive breast cancer presents negative correlation. But under negative condition of HER-2/neu, the tumor grading of invasive breast cancer shows positive correlation.

目錄
中文摘要	I
英文摘要 (ABSTRACT IN ENGLISH)	II
縮 寫 表	V
表目錄	VIII
圖目錄	X
第一章	緒論	1
1.1	乳癌在台灣	1
1.2	組織晶片之介紹	1
1.3	ER/PR、HER-2/NEU之介紹	2
1.4	NOTCH PATHWAY的介紹	4
1.5	研究動機	5
第二章	實驗材料與方法	6
2.1	研究標本篩選	6
2.2	組織晶片製備	6
2.3	H&E染色	8
2.4	免疫組織化學染色(IMMUNOHISTOCHEMISTRY, IHC)	9
2.5	TMA中IHC染色結果評估方法	12
2.6	TMA統計分析方法	13
第三章	實驗結果	14
3.1	樣本分佈之分析	14
3.2	NOTCH1免疫組織化學染色之分析	14
3.3	JAGGED1 免疫組織化學染色之分析	15
3.4	DELTA 免疫組織化學染色之分析	16
3.5	所有樣本的免疫組織化學染色之分析	17
第四章	討論	19
第五章	結論與展望	22
參考文獻	23
圖表	26
附錄	52
 
表目錄
表一 .  組織晶片、基因晶片、蛋白質晶片區別比較	26
表二 .  Notch訊息組成要件表	27
表三 .  本研究之樣本表現乳腺癌病人年紀和組織特性之分佈	28
表四 .  本研究樣本之病人，乳腺癌腫瘤蛋白ER、HER-2/neu之分佈統計	29
表五 .  本研究樣本之病人，乳腺癌腫瘤蛋白PR、HER-2/neu之分佈統計	29
表六 .  本研究樣本乳腺癌病人年齡分佈範圍概況	30
表七 .  本研究樣本乳腺癌病人癌組織解剖部位之分佈概況	30
表八 .  Spearman's rank correlation coefficient 。HER-2/neu陽性時，Notch1與ER/PR、惡性程度(grade)、HER-2/neu四者相關性分析	31
表九 .  Spearman's rank correlation coefficient 。ER/PR陽性而HER-2/neu陰性時，Notch1與ER/PR、惡性程度(grade)、HER-2/neu四者相關性分析	31
表十.  Spearman's rank correlation coefficient 。HER-2/neu陽性條件下，對於Jagged1表現分別與ER/PR、惡性程度(grade)、HER-2/neu四者及上述Notch1、Delta相關性分析	32
表十一 .  Spearman's rank correlation coefficient。ER/PR陽性及HER-2/neu陰性條件下，對於Jagged1表現分別與ER/PR、惡性程度(grade)、HER-2/neu四者及上述Notch1、Delta相關性統計分析表	33
表十二 .  Spearman's rank correlation coefficient 。HER-2/neu陽性時，Delta與ER/PR、惡性程度(grade)、HER-2/neu四者相關性分析	33
表十三 .  Spearman's rank correlation coefficient 。HER-2/neu陰性時，Delta與PR、惡性程度(grade)、HER-2/neu四者相關性分析	34
表十四 .  Spearman's rank correlation coefficient。五十九例乳腺癌組織統計分析結果表	35
表十五 .  Notch1和Jagged1在Breast Cancer中表現的相關性	36
表十六 .  Notch1和Delta在Breast Cancer中表現的相關性	36
表十七 .  Delta和Jagged1在Breast Cancer中表現的相關性	36

 
圖目錄
圖一 .  Notch 訊息傳遞路徑	37
圖二 .  乳腺癌組織惡性程度系統(NOTTINGHAM GRADING SYSTEM)	38
圖三 .  實驗流程圖	39
圖四 .  建立組織陣列流程圖。a：定位取樣 b：置入樣本 c：切片，完成晶片製作	40
圖五 .  A:乳腺癌TMA之H&E染色。	41
圖五(續) .  B:呈現high grade carcinoma高惡性度組織相 (200×)。C:呈現low grade carcinoma低惡性度組織相 (200×)。	42
圖六 .  本研究樣本之病人，乳腺癌腫瘤蛋白HER-2/neu、ER之分佈概況	43
圖七 .  本研究樣本之病人，乳腺癌腫瘤蛋白HER-2/neu、PR之分佈概況	43
圖八 .  A:為乳腺癌TMA上一個樣本組織相，H&E染色 (100×)。而B:為同一標本之Notch1染色，呈現為陽性 (400×)。	44
圖八(續) .  C:為另一乳腺癌TMA上的樣本組織相，H&E染色 (100×)。而D:為同一標本之Notch1染色，呈現為陰性 (400×)。	45
圖九 .  A、B為同一組織而C、D為另一相同組織。A：乳腺癌TMA之H&E染色 (200×)。B：在HER-2/neu陽性的條件下，Notch1陰性，呈現high grade carcinoma高惡性度組織相 (200×)。	46
圖九(續) .  C為乳腺癌TMA之H&E染色 (200×)。D為Notch1染色，呈現陽性，為low grade carcinoma低惡性度組織相 (200×)。	47
圖十.  為Jagged1染色，呈現為陽性 (400×)。	48
圖十一 .  為Delta染色，呈現為陽性 (400×)。	49
圖十二 .  在乳腺癌組織中，Delta染色位置在細胞質呈現陽性 (400×)。	50
圖十三 .  A：為IHC染色強度次數分佈柱狀圖。B：為IHC腫瘤細胞染色所佔百分比次數分佈柱狀圖。	51

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