HER-2/neu amplification and expression in papillary lesion of breast

Abstract

根據行政院衛生署統計資料，乳癌已成為國內女性癌症發生率的第一位，死亡率為第四位，已取代子宮頸癌成為國內婦女癌症發生率首位，因此乳房篩檢也成為我國婦女預防保健的重要課題。隨著婦女篩檢參與率的增長，早期乳房病灶的發現率也隨之提昇，同時增加了乳房的組織病理切片率。而乳房疾病之一類的乳突樣病灶中，其範圍涵蓋了良性的乳突瘤到侵襲性的乳突樣癌，各種乳突樣病灶在組織切片的型態特徵上均很相似，不易鑑別，經常造成病理醫師在判讀上之困擾。人類表皮生長因子受體II（HER-2/neu），是原癌基因，可加劇乳腺癌之癌化，其參與細胞生長和分化的訊息傳遞，這種受體的過度表現在乳腺癌，能增加疾病的復發而且具有更差的預後。 目前在基因體學、附基因體學及蛋白質體學各層面的研究皆積極設法開發具有預防、診斷鑑別力的生物標記，並期盼作為新興治療或藥物開發的研究對象。本研究主要採行以醫院為基礎的病例世代研究方式，由三軍總醫院病理部資料庫中進行17年的回溯性研究探討，挑選診斷為乳房乳突樣病灶之303例檢體進行分析研究。 本實驗利用于承平老師實驗室所建立之乳房疾病組織微陣列，將乳房乳突樣病灶以螢光原位雜交法偵測HER-2/neu基因擴增之情形， 以及以免疫組織化學染色法檢測HER-2/neu蛋白質之表現情形，比較乳突樣病灶從良性到惡性之進程中HER-2/neu的動態演變，以深入探討其可能的分子病理機制。 我們的實驗結果顯示，HER-2/neu基因之擴增與蛋白質之表現，確實有助於鑑別良性和惡性的乳房乳突樣病灶，且HER-2/neu蛋白質之表現同時具有癌化風險預測的臨床應用價值。故HER-2/neu免疫組織化學染色法可以用為組織病理鑑別診斷之利器，可以提供病患適當的醫療監測，進而適時掌握對乳癌診治之黃金時刻。According to the statistic data from Department of Health of Taiwan, breast cancer has become the first one in cancer incidence and the fourth place in cancer mortality in Taiwanese women. Screening of breast cancer has become an important issue in preventive medicine in Taiwan. Due to the increment of breast cancer screening rate, many early breast lesions have been identified which causes a raise of breast biopsy compared with the previous. The papillary lesion is an entity of breast ranging from benign intraductal papilloma to invasive papillary carcinoma and all of them are histopathologically similar which makes pathologists hard to distinguish them. Human epidermal growth factor receptor II (HER-2/neu) is a proto-oncogene, and it can exacerbate breast cancer progression. HER-2/neu signal transduction promotes cell proliferation and differentiation. HER-2/neu overexpression in breast cancer has been associated with more recurrence and worse prognosis. Recent studies of genomics, epigenetics and proteomics are devoting to develop biomarkers for diagnosis and outcome prediction, and also looking forward to drug development. Here, we adopted a hospital-based case-cohort study and use the breast disease tissue microarray established by Dr. Cheng-Ping Yu to investigate the papillary lesions of the breast including 303 cases. In this study, we performed fluorescence in situ hybridization and immunohistochemistry to investigate the gene amplification and protein expression of HER-2/neu then analyze the role as well as molecular mechanism of HER-2/neu in carcinogenesis of breast papillary lesions. Our results show that immunohistochemistry of fluorescence in situ hybridization of HER-2/neu gene amplification and HER-2/neu protein expression both help to distinguish benign and malignant papillary lesions of breast. In addition, immunohistochemistry of HER-2/neu protein expression has a role in cancer risk prediction. We suggest that immunohistochemistry of HER-2/neu protein expression could be a useful biomarker and method for the differential diagnosis of papillary lesions in breast biopsy which can provide good patient monitoring and treatment timing of breast cancer clinically.中文摘要··································································································iii 英文摘要··································································································v 第一章、前言··························································································01 第一節 乳癌之流行病學····································································01 第二節 乳房組織結構及疾病概述····················································01 第三節 乳房乳突樣病灶(papillary lesions)之分類及其臨床生物特性·······························································································05 第四節 HER-2/neu基因的介紹·························································10 第五節 研究動機················································································13 第二章、研究目的··················································································14 第三章、材料與方法··············································································16 第一節 檢體資料收集········································································16 第二節 建構組織微矩陣(tissue microarray)······································18 第三節 螢光原位雜交法(Fluorescence in situ Hybridization, FISH)20 第四節 免疫組織化學染色(immunohistochemistry, IHC)················30 第五節 統計分析················································································32 第四章、結果··························································································34 第一節 乳房乳突樣病灶與良性乳房疾病罹癌風險比值之比較·····34 第二節 HER-2/neu基因擴增、HER-2/neu蛋白表現以及其他乳癌危險因子在良、惡性乳突樣病灶中的表現差異····················37 第三節 HER-2/neu基因擴增與否、HER-2/neu蛋白表現高低與良性乳突樣病灶之乳癌風險評估················································38 第四節 HER-2/neu基因擴增與否、HER-2/neu蛋白表現高低在組織病理診斷之輔助角色····························································40 第五章、討論··························································································42 第一節 利用台灣、美國兩大醫學中心探討良性乳突樣病灶癌化的風險差異····················································································42 第二節 HER-2/neu基因擴增或蛋白高表現均提高乳癌風險·········44 第三節 HER-2/neu基因擴增及蛋白表現與乳房乳突樣病灶之惡性程度相關····················································································45 第四節 HER-2/neu基因擴增及蛋白表現可用以輔助良、惡性乳突病灶之組織病理診斷································································47 第六章、結論··························································································51 第七章、參考文獻·················································52 圖表目錄 表1. 良性乳突樣病灶與其他良性乳房疾病之乳癌風險比·················57 表2. 合併良性乳突樣病灶與其他良性乳房疾病之乳癌風險比·········57 表3. HER-2/neu基因擴增及蛋白表現與乳癌流行病學危險因子在良、惡性乳突樣病灶中之差異性························································58 表4-1. HER-2/neu基因擴增與乳房乳突樣病灶之乳癌風險評估·······59 表4-2. HER-2/neu蛋白表現與乳房乳突樣病灶之乳癌風險評估·······59 表5-1. HER-2/neu基因擴增與否在不同乳房病理組織中的分佈情形 60 表5-2. HER-2/neu蛋白表現高低在不同乳房病理組織中的分佈情形 60 圖1. H&E染色下各種乳房乳突病灶之組織病理特徵·····················61 圖2. 螢光原位雜交下，乳房乳突病灶之HER-2/neu基因擴增情形·62 圖3. 免疫組織化學染色下，乳房乳突病灶之HER-2/neu蛋白質表現情形····································································································6