臺灣博碩士論文加值系統

在西元2003年全年台灣地區癌症死亡原因當中，胰臟癌在男性中排名第九位，女性排名第八位，是一種不容忽視的癌症。為了發展新的診斷、治療診斷及預防方式，確認胰臟癌的多步驟腫瘤形成過程成為當前重要的課題，但是到目前為止我們對涉及胰臟癌的基因所知有限。
Hruban等人(2000)創立胰管腺癌形成之進展模式(genetic progression model for pancreatic intraepithelial neoplasias)，其中牽涉到一系列多種基因的改變。然而因胰臟癌複雜的基因改變及部分染色體區域的套數增多或減少，難以全盤性地用一般的方法鑑定。因此吾人在此研究當中，利用競爭性基因體核酸雜交法(comparative genomic hybridization, CGH)全盤性地瞭解胰臟癌組織中基因體染色體的變異區段，並試圖瞭解其中所涉及之抑癌或致癌基因。
在此研究中所蒐集的23例胰臟癌檢體當中，16例為胰管腺癌(pancreatic ductal adenocarcinoma)，4例為內分泌性胰臟癌(endocrine carcinoma)，1例為腺泡細胞癌(acinar cell carcinoma)，和2例黏液性腺癌(mucinous carcinoma)。吾人利用CGH分析23例胰臟癌檢體，結果發現腫瘤基因變異情形如下：在染色體13q(佔61%)、7p、8q、和14p(以上各佔57%)、5q、13p、19p和21p(以上各佔52%)、1q、5p、6p和11p(以上各佔48%)出現染色體套數增加的情形；在染色體16q、18q (26%)和1p (22%)出現套數減少的情形，由此可知腫瘤的形成和上述區段染色體的變異可能有所關聯。在16例胰管腺癌中有10例(佔63%)在染色體8q出現套數增加的情況，各有9例(56%)在染色體13q和14p，8例(50%)在染色體1q、5q、7p、11q、13p和21p出現相同的情形。而5例(31%)在染色體18q則出現套數減少的情形，4例(25%)在染色體16q出現相同的情形。而在4例內分泌性胰臟癌中，其中三例出現染色體6q套數增加的情形。由此推知胰臟腫瘤的形成和染色體的變異有其關聯性。綜觀各例之結果，發現在於染色體8q套數增加與胰管腺癌之形成有強烈之相關性；而6q套數增加則與內分泌性胰臟癌之衍生有特殊之意義。此一研究如在將來進一步以微陣列競爭性基因體核酸雜交法(array CGH)，則可提升其解析度，將可針對特定染色體段落，乃至其基因之套數加以分析，瞭解在各個階段之胰臟癌的演化與特定基因之關連性，進而試圖瞭解特定種類胰臟癌之癌化過程之相關機轉。

The mortality of pancreatic cancer in male is the ninth leading cause of cancer death during year 2003 in Taiwan, and that in female ranks eighth. In order to develop the new diagnostic, therapeutic and preventive methods of the disease, we ought to define the progression of the tumorigenesis mechanism of the cancer. However, what we know about the genetic mechanism of the disease is not enough.
Hruban et al. (2000) proposed the genetic progression model for pancreatic intraepithelial neoplasias. It involved a set of genetic variations. The complicated copy number amplification and deletion in particular regions of chromosomes were difficult to detect with usual methods. Therefore, we used comparative genomic hybridization (CGH) to define the variant regions of chromosomes in pancreatic cancers, and tried to define the tumor suppressor genes and oncogenes involved.
Twenty-three cases of pancreatic cancer were collected, including 16 cases of pancreatic ductal adenocarcinoma, 4 cases of endocrine carcinoma, a case of acinar cell carcinoma, and 2 cases of mucinous adenocarcinoma. After CGH analyses to the 23 cases the copy number abnormality showed that the gain occured in 14 cases (61%) at chromosomes 13q, in 13 cases (57%) at each of 7p, 8q and 14p, in 12 cases (52%) at each of 5q, 13p, 19p and 21p, in 11 cases (48%) at each of 1q, 5p, 6p and 11p, and the loss in 6 cases (26%) at chromosome 16q and 18q, and in 5 cases (22%) at 1p. We also noticed among 16 cases of pancreatic ductal adenocarcinoma, that the copy number gain occurred in chromosomes 8q (63%), 13q, 14p (56%), 1q, 5q, 7p, 11q, 13p and 21p (50%), and loss in chromosomes 18q (31%) and 16q (25%). Also, in 3 among 4 cases of endocrine carcinoma it showed the gain at chromosome 6q. The results imply that the particular type of tumor may be related to the certain genomic aberration in the identified specific regions. Further investigation with the array CGH, which increases the resolution in detecting the particular regions of chromosomes, may facilitate the understanding of the specific genes that may be involved. It can be then analyzed in correspondence with the various stages of the tumors, and the tumorigenesis progression of pancreatic carcinoma.

目錄
指導教授推薦書………………………………………………
口試委員會審定書……………………………………………
電子檔案上網授權書………………………………………… iii
紙本論文著作授權書………………………………………… iv
誌謝…………………………………………………………… v
中文摘要……………………………………………………… vi
英文摘要……………………………………………………… viii

第一章 緒論……………………………………………….. 1
1.1 胰臟介紹……………………………………………… 1
1.2 胰臟癌的流行病學…………………………………… 2
1.3 胰臟癌的種類………………………………………… 3
1.4 胰臟癌臨床症狀……………………………………… 4
1.5 胰臟癌分期…………………………………………… 5
1.6 胰臟癌的診斷與治療………………………………… 6
1.7 胰臟癌的研究………………………………………… 8
第二章 實驗設計………………………………………….. 14
第三章 材料與方法……………………………………….. 15
3.1 檢體的取得…………………………………………… 15
3.2 組織切片、染色……………………………………… 15
3.3 DNA萃取及DNA探針的製備………………..… 16
3.4 正常人類中期淋巴球染色體之製備…………………. 17
3.5 競爭性基因組核酸雜交……………………………… 18
3.6 影像分析……………………………………………… 20
3.7 陽性控制組之腫瘤細胞之培養…………………… 21
3.8 轉移細胞……………………………………………… 22
3.9 培養細胞之DNA萃取………………………………. 23
第四章 結果………………………………………………… 24
4.1 胰臟癌病例報告………………………………………. 24
4.2 23例胰臟癌之CGH之染色體變異分析……………. 24
4.3 16例胰管腺癌之CGH之染色體變異分析…………. 25
4.4 各病例所屬之胰臟癌型態及染色體變異情形………. 25
4.5 胰臟癌分期與染色體之變異分析…………………….25
第五章 討論………………………………………………… 27
圖表及說明…………………………………………………… 34
附錄…………………………………………………………… 65
參考文獻……………………………………………………… 73

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