Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/13336
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dc.contributor.advisor王孟亮zh_TW
dc.contributor.advisor張仕杰zh_TW
dc.contributor.author陳信帆zh_TW
dc.contributor.authorCHEN, HSIN-FANen_US
dc.date2005zh_TW
dc.date.accessioned2014-06-06T06:50:40Z-
dc.date.available2014-06-06T06:50:40Z-
dc.identifier.urihttp://hdl.handle.net/11455/13336-
dc.description.abstract血紅素為傳統的globin家族成員,儲存於紅血球之中,負責運輸氧氣。在人與犬的乳腺腫瘤組織與犬正常乳腺組織中,曾經發現有血紅素蛋白的存在。血紅素已經被證實能夠刺激腫瘤細胞分泌基質金屬蛋白酵素和吸附腫瘤細胞,以促進腫瘤細胞的增殖或轉移。因此,腫瘤組織中血紅素的表現與腫瘤發展有所關聯。 本研究在犬隻的正常乳腺與乳腺腫瘤組織的SDS-PAGE蛋白電泳圖中發現,於15 kD的位置有不同表現量的蛋白條帶出現,其表現量以正常乳腺最強、惡性乳腺腫瘤次之、良性乳腺腫瘤最弱。另外,在乳腺腫瘤分期上的表現量,則隨著分期的增加,15 kD蛋白的表現量有上升的趨勢,但是在第五期乳腺腫瘤的表現則又下降。經由Western blotting分析,發現正常乳腺所表現出相當多的15 kD蛋白為hemoglobin。進一步以質譜儀分析此蛋白條帶,確定15 kD蛋白內含有hemoglobin α chain與β chain。另外,用正常乳腺組織以Western blotting偵測glycophorin A,結果發現並不帶有紅血球細胞膜上的特徵蛋白質glycophorin A,這表示我們所偵測到的hemoglobin並非來自血液中紅血球。目前關於犬隻正常乳腺或乳腺腫瘤hemoglobin表現的機制與功用仍未知,仍需更進ㄧ步的研究。zh_TW
dc.description.abstractHemoglobin is a classical member of the globin family and is stored in red blood cells. Hemoglobin is a typical protein for transporting oxygen through the body. Besides canine normal mammary gland, hemoglobin has been detected in both human and canine mammary gland tumor. Some reports showed that hemoglobin can up-regulate metalloproteinase-9 and help tumor cell's adhesion. Therefore, hemoglobin may play a role in tumor progression. We find that there is a prominent band with different degree of expression at approximately 15 kD in SDS-PAGE in canine normal and in neoplastic mammary gland. It has the strongest expression in normal mammary gland tissue, and follow by the malignant and the benign mammary gland tumor. In addition, the expression of the 15 kD protein increase with the clinical stage serious level along. Then, its expression falls at clinical stage V. After Western blotting analysis, the 15 kD protein in normal mammary gland is shown to be hemoglobin. Furthermore, mass spectrometry results demonstrated that the 15 kD protein is hemoglobin α chain and β chain. We also use Western blotting to detect a characteristic red blood cell membrane protein, glycophorin A, in the normal mammary gland tissue, and we find glycophorin A is not detected in the normal mammary gland tissue. Thus, it indicated the hemoglobin we found is not from the red blood cells. The mechanism and the function of hemoglobin expressed in the normal or neoplastic mammary gland was not known yet.en_US
dc.description.tableofcontents目錄 目錄...................................................................................................................Ⅰ 圖次...................................................................................................................Ⅲ 表次...................................................................................................................Ⅳ 中文摘要...........................................................................................................Ⅴ 英文摘要...........................................................................................................Ⅵ 第一章 緒言.................................................................................................1 第二章 文獻探討.........................................................................................2 第一節 乳腺腫瘤之探討.........................................................................2 1-1 犬乳腺腫瘤之流行病學............................................................2 1-2 乳腺腫瘤病因學........................................................................2 1-3 犬乳腺腫瘤的種類、組織病理學分類和生物表現................4 1-4 犬乳腺腫瘤的臨床表現............................................................6 1-5 犬乳腺腫瘤的分期....................................................................7 1-6 犬乳腺腫瘤的預後因子............................................................9 1-7 乳腺腫瘤中血管新生現象......................................................10 第二節 血紅素表現之探討...................................................................11 2-1 血紅素基因之介紹..................................................................11 2-2 血紅素蛋白胺基酸序列..........................................................13 2-3 血紅素之功能..........................................................................13 2-4 血紅素在乳腺腫瘤組織的表現..............................................17 2-5 血紅素在腫瘤組織中可能扮演的角色..................................17 第三節 紅血球細胞膜特徵蛋白glycophorin A之介紹......................20 第三章 材料與方法...................................................................................21 第一節 樣本的採樣...............................................................................21 1-1 犬隻正常乳腺、乳腺腫瘤與其餘正常組織的採樣..............21 1-2 血液樣本的取得......................................................................25 第二節 利用SDS-PAGE進行蛋白質電泳分析..................................25 2-1 乳劑之製作..............................................................................25 2-2 蛋白質定量分析......................................................................26 2-3 SDS-PAGE之製備..................................................................27 2-4 SDS-PAGE蛋白質電泳條帶分析..........................................28 第三節 利用西方轉漬法偵測血紅素蛋白...........................................28 第四節 利用西方轉漬法偵測glycophorin A蛋白..............................30 4-1 純化血液中紅血球細胞膜......................................................30 4-2 西方轉漬法..............................................................................30 第五節 質譜儀分析15 kD蛋白條帶...................................................30 第四章 結果...............................................................................................31 第一節 實驗樣本資料與組織學診斷...................................................31 第二節 利用SDS-PAGE進行犬組織蛋白質電泳分析......................31 第三節 利用西方轉漬法偵測血紅素蛋白...........................................32 第四節 利用西方轉漬法偵測glycophorin A蛋白..............................32 第五節 質譜儀分析15 kD蛋白條帶...................................................32 第五章 討論...............................................................................................45 參考文獻...........................................................................................................48 附錄...................................................................................................................55 圖次 圖1、血紅素基因結構示意圖.........................................................................12 圖2、犬隻與人的α-globin胺基酸序列對照................................................14 圖3、犬隻與人的β-globin胺基酸序列對照................................................14 圖4、血紅素蛋白結構示意圖.........................................................................15 圖5、血紅素與氧分子結合示意圖.................................................................15 圖6、犬隻乳腺的正常組織與腫瘤組織的SDS-PAGE蛋白質電泳............19 圖7、犬正常乳腺組織與乳腺腫瘤SDS-PAGE電泳分析............................34 圖8、犬正常乳腺組織與乳腺腫瘤SDS-PAGE電泳分析............................35 圖9、犬正常乳腺組織、乳腺腫瘤與淋巴結SDS-PAGE電泳分析............36 圖10、正常乳腺組織、其餘正常組織與血液SDS-PAGE電泳分析..........37 圖11、以組織學分組,15 kD蛋白條帶的平均表現量比較........................38 圖12、以乳腺腫瘤分期,15 kD蛋白條帶的平均表現量比較....................39 圖13、利用西方轉漬法進行血紅素蛋白電泳分析.......................................40 圖14、利用西方轉漬法進行glycophorin A蛋白電泳分析..........................41 圖15、正常乳腺組織與乳腺腫瘤組織SDS-PAGE電泳分析......................42 圖16、圖15 Lane 1正常乳腺表現之15 kD蛋白條帶經質譜儀分析結果..43 圖17、圖15 Lane 1 正常乳腺表現之15 kD蛋白條帶經質譜儀分析結果..43 圖18、圖15 Lane 5 第四期惡性乳腺腫瘤表現之15 kD蛋白條帶經質譜儀 分析結果.............................................................................................44 圖19、圖15 Lane 5 第四期惡性乳腺腫瘤表現之15 kD蛋白條帶經質譜儀 分析結果.............................................................................................44 表次 表1、世界衛生組織之犬乳腺腫瘤的組織病理學分類...............................5 表2、犬乳腺腫瘤分期依據世界衛生組織的TNM系統............................8 表3、犬乳腺腫瘤的臨床分期.......................................................................8 表4、血紅素片段的生物活性.....................................................................16 表5、 實驗樣本資料...................................................................... ..........22zh_TW
dc.language.isoen_USzh_TW
dc.publisher獸醫學系zh_TW
dc.subjecthemoglobinen_US
dc.subject血紅素zh_TW
dc.title血紅素在正常乳腺或乳腺腫瘤表現之研究zh_TW
dc.titleExpression of hemoglobin in the normal or neoplastic mammary glanden_US
dc.typeThesis and Dissertationzh_TW
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