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研究生:高韶蔭
研究生(外文):Kao,Shau-Yin
論文名稱:在胃癌中matriptase的活化及其機轉
論文名稱(外文):The activation of matriptase and the possible mechanism in gastric cancer
指導教授:王正康
學位類別:碩士
校院名稱:國防醫學院
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:91
中文關鍵詞:胃癌
外文關鍵詞:matriptase
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Matriptase屬於第二類跨膜型絲胺酸蛋白酶 (transmembrane serine protease),主要廣泛分布於各個組織的上皮細胞中。Matriptase在細胞中新生成時為不具活性的單鏈型蛋白酶,再經由S1P (sphingosine-1-phosphate) 的作用,形成具活性的雙鏈型蛋白酶,活化後的matriptase會立即與 HAI-1 形成可逆性的結合,並藉此調控matriptase的活性。而已知此接受體可能為G protein coupled receptor當中的EDG-3 (Endothelial Differentiation Gene),但詳細的訊息傳遞機制仍不清楚。由過去的臨床研究顯示,在一些上皮細胞癌中包括:乳癌、子宮頸癌、大腸癌中matriptase有過度的表現,進一步的知道在卵巢癌及前列腺癌中則是matriptase/HAI-1的比例增加,且隨著轉移程度的增加,有更多的表現。胃癌在台灣為高發生率的疾病,且大部分都屬於胃腺癌,過去卻鮮少探討matriptase與胃癌的關係。我們與三總一般外科陳正榮醫師合作,將取得的胃癌檢體,利用冷凍切片方式並進行免疫組織化學染色,發現在胃癌組織中matriptase的確有大量的表現,並伴隨HAI-1的表現,若依不同的淋巴結的侵犯程度、癌細胞的侵犯程度、性別、年齡層及細胞的分化情形比較,發現隨著淋巴結受侵犯增加、癌細胞的侵犯程度增加、年齡的增加,可以看到活化態matriptase的比例都有增加的現象,更觀察到隨著臨床分期的增加,可以偵測到活化態matriptase有明顯表現的比例亦增加了,在早期胃癌中可以偵測到活化態matriptase的比例只佔33.3%;但在後期則比例提高了許多甚至達100%。此外在女性中可以看到活化態matriptase表現的比例較男性高;然而在細胞分化的部分,可能因檢體數太少,目前還無法觀察到明顯的表徵。另外利用EDG-3的抗體,偵測EDG-3的表現,可是並無很明確的結果,可能還需要累積更多的檢體,才能進一步探討其在胃癌中可能的機制。
Matriptase is a type II transmembrane serine protease, which is ubiquitously expressed in the epithelial tissues. After activation, activated matriptase is bound by HAI-1 immediately, and become a dynamic complex. The activation mechanism of matriptase could be driven by S1P. S1P may act as a ligand of EDG3, a G protein coupled receptor, and activate matriptase through unknown signaling pathway. Previous studies suggested that matriptase is overexpressed in several epithelial cancers, including breast cancer, cervical cancer, and colon cancer. Furthermore, the matriptase : HAI-1 ratio is increased in ovarian and prostate cancer of more advanced stage. In Taiwan, gastric cancer is the a prevalence cancer disease and most of them are adenocarcinoma. In the past studies, the relationship of matriptase and gastric cancer has not yet been well identified. We obtained specimens from the department of general surgery, TSGH, and utilized these specimens to study the correlation between matriptase and gastric cancer. We found that matriptase along with HAI-1 are largely expressed in gastric cancer by using frozen section and IHC methods. We also analyzed the influence of lymph node invasion, differentiation status, gender, age, and different invasion of tumor on matriptase activation in gastric cancer. Our results demonstrated strong correlation between cancer stage and matriptase activation. Active matriptase can be detectd in all late stage gastric cancer but only 33.3% in early stage. We also found more active matriptase in female, however the specimen amount in female is less than male. As for age and lymph node invasion are positively correlated with matriptase activation. However we could not find any relationship between matriptase activation and differentiation. In addition, the EDG3 expression in gastric cancer was also examined by using EDG3 antibody but we could not get significant results. We still need to perform some further experiments to verify the role of EDG3 in gastric cancer.
目錄 I
表及圖目錄 IV
縮寫表 VI
中文摘要 VII
英文摘要 IX
第一章 緒論 1
第一節 蛋白酶(Proteases) 1
第二節 Matriptase 2
壹、Matriptase的發現與基本介紹 2
貳、Matriptase的表現與活化 3
第三節 HAI-1的結構與功能 6
壹、HAI-1的結構 6
貳、HAI-1的功能 7
参、HAI-1在調控matriptase 活化的過程中所扮演的角色 7
第四節 EDG家族 9
第五節 Matriptase可能扮演的生理角色 11
第六節 在不同癌症中matriptase和HAI-1 的異常表現 14
壹、matriptase/HAI-1與乳癌的關係 14
貳、matriptase/HAI-1與子宮頸癌的關係 14
参、matriptase/HAI-1與卵巢癌的關係 15
肆、matriptase/HAI-1與前列腺癌的關係 16
伍、matriptase只有在上皮來源(epithelial origin)的腫瘤可以看到表現 16
第七節 胃臟及胃癌的流行病學(epidemiology)與分類 17
壹、胃的基本構造與組成 17
貳、胃癌的流行病學(epidemiology)與致病因子 18
参、胃癌的診斷與分期 20
第八節 實驗目的 22
第二章 實驗材料與方法 24
實驗材料 24
一、藥品試劑 24
二、實驗儀器 26
三、緩衝液及SDS-PAGE之備製 27
四、病人檢體(specimen) 之取得與保存 29
實驗方法 29
一、冷凍切片 29
二、免疫組織化學染色 31
三、蘇木紫-伊紅染色 (Hematoxylin & Eosin stain) 32
第三章 結果 34
第一節 Matriptase 在癌細胞與鄰近正常細胞中的表現 34
壹、Total matriptase的表現 34
貳、活化態(active) matriptase的表現 35
第二節 活化態matriptase在男性與女性中的表現 35
第三節 比較不同年齡層中活化態matriptase的表現 36
第四節 觀察癌細胞是否侵犯淋巴結(N)時活化態matriptase的表現 37
第五節 觀察癌細胞侵犯至不同層(T)時活化態matriptase的表現 38
第六節 比較不同分化程度中活化態matriptase的表現 39
第七節 依不同的臨床診斷期別比較活化態matriptase有表現的比例 39
第八節 活化態matriptase和EDG3在胃癌細胞中的表現 40
第四章 討論 42
第一節 胃癌中matriptase、活化態matriptase及HAI-1間的表現 43
第二節 活化態matriptase的表現可能和胃癌的臨床分期有關 44
第三節 胃癌中,活化態matriptase的表現可能與性別及荷爾蒙有關 46
第四節 胃癌中,活化態matriptase和EDG3間的關係 48
第五節 未來的工作與方向 49
參考文獻 51
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