臺灣博碩士論文加值系統

摘要
促胃液分泌激素（gastrin）是一荷爾蒙，由胃幽門腔上的G細胞所分泌。主要有兩個功能，一者可促進胃壁細胞（parietal cell）分泌胃酸；二來為一生長因子（growth factor）可誘發胃黏膜細胞的增生。 近來有報告指出gastrin的過量分泌會造成腸胃道癌症或非腸胃道腫瘤，如胰臟癌(pancreatic carcinoma)、肺小細胞癌(small cell lung cancer)、腎臟癌(renal tumor)的產生。此外，由胃酸分泌過多所造成的消化性潰瘍，亦跟gastrin有間接的關係。本論文主要目的為發展一個可誘導動物體內產生抗gastrin抗體的抗原，利用抗體專一性中和gastrin的能力，期望其可成為一安全、有效治療腫瘤或潰瘍之治療性疫苗。我們使用LAE技術（線性重複抗體辨認區段技術）來製作具有多數重複之gastrin抗原決定基（gastrin epitope）的DNA片段，將其銜接至蛋白質表現載體中。經大腸桿菌表現後，得到包含綠膿桿菌外毒素A之受體結合器（the receptor binding domain of Pseudomonas aeruginosa exotoxin A）與多數重複之gastrin抗原決定基的抗原。將此抗原免疫實驗動物後，經酵素免疫分析，證實此抗原可誘導動物體產生抗gastrin抗體。在AR42J細胞實驗中，藉由觀察ICER基因的表現，確認抗gastrin抗體可抑制gastrin與其受體的結合。綜上所述，利用LAE技術所製作的gastrin抗原，的確可在動物體內誘導產生抗gastrin抗體的產生；且此抗體在細胞層次具有拮抗gastrin的生物功能。然而此抗原是否能在動物體內治療gastrin相關疾病，則有待進一步的研究發現。

Abstract
Gastrin, produced by G cells in the gastric antrum, has been identified as the circulation hormone responsible for stimulation of acid secretion from the parietal cell. It also has an important trophic of growth —promoting influence on the gastric mucosa. Recent studies showed that excessive secretion of gastrin promotes the growth of some gastrointestinal cancers, as well as non-gastrointenstinal cancers, including pancreatic carcinoma, small cell lung cancer, and renal tumors. Peptic ulcers caused by increased stomach acid which is also dependent on gastrin. In this study, we propose to develop immunogens which can induce anti-gastrin antibody and neutralize overexpressed gastrin, and hopefully they could be used as safe and efficient therapy vaccine. Toward this end, we used Linear-Array Epitope (LAE) techniques to construct DNA fragments encoding multicopies of gastrin epitopes, followed by subcloning it into a protein expression vector. The immunogen is a fusion protein containing the receptor binding domain of Pseudomonas aeruginosa exotoxin A and multicopies of gastrin epitopes. Immunizing with the immunogens in rabbits could induce the anti-gastrin antibodies responses. On AR42J cells, through the detection of the suppression of ICER gene expression, we identify anti-gastrin antibody could inhibit the interaction between gastrin and gastrin receptor. In conclusion, anti-gastrin antibodies induced by gastrin immunogens could effectively neutralize gastrin activity in vitro and their therapeutic effects in vivo are under investigation now. The study also may pave the way to the development of anti-gastrin vaccine for gastrin dependent diseases.

目錄
頁次
中文摘要………………………………………1
英文摘要………………………………………2
緒論……………………………………………3
材料與方法…………………………………..12
結果…………………………………………..26
討論…………………………………………..34
參考文獻……………………………………..38
附圖…………………………………………..43
附錄…………………………………………..63
試劑配方……………………………………..66

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