臺灣博碩士論文加值系統

神經膠質瘤源於神經膠細胞，其數量占大腦細胞的百分之九十，功能為分泌神經成長因子誘發神經纖維生長、形成髓鞘及血腦屏障。最常見的神經膠質瘤為神經膠母細胞瘤，神經膠質瘤形成的因素至今尚未完全明朗。由於神經膠質瘤生長邊界不明確，以手術方式無法清除乾淨，對於化學治療及放射線治療具有高度的抵抗性而不易治療。另一方面，因為神經膠質瘤的高度入侵性及致死率，使得科學家們致力於去開發及尋找新的治療方法及治療標的基因。之前研究發現同時給予interlukin-2 (IL-2)及組織胺治療能有效地抑制惡性神經膠質瘤的生長及血管新生，這也暗示著組織胺有抑制神經膠質瘤生長的功能。組織胺在細胞內主要藉由histamine-N-methyltransferase (HNMT)來代謝。前人的實驗結果已經證實組織胺含量及其代謝物和癌症有著一定的相關性，然而卻沒有相關的文獻直接指出HNMT和神經膠質瘤的關係或探討HNMT相關的致病機轉，本論文發現不論是神經膠質瘤組織或細胞株，HNMT的表現量都較正常腦組織多，高度表現的HNMT暗示著抗癌的組織胺含量下降。另一方面，神經膠質瘤促進因子之一的epithelial growth factor (EGF)會增加HNMT的表現及HNMT在細胞中位置的轉移。針對致命的神經膠質瘤，我們的結果提供HNMT是另一個新的治療標的。

Giloma derive from glial cell, which contribute about 90% of brain cells. The functions of glial cells are offering support and nutrition, forming myelin sheath and blood brain barrier.The most common glioma is glioblastoma multiforme (GBM) and the possible causes are not fully understood. Neurosurgery unable to remove the glioma from normal brain clearly；Glioma even resisten to high dose of radiotherapy and chemotherapy. The prognosis of malignant glioma remains dismal and the estimated median survival time is <1 year.It is emergent to develop new therapic policy and find out target genes. Previous data investigate that cotreatment with interleukin-2 (IL-2) and histamine inhibits growth and angiogenesis in malignant glioma. It also suggests that histamine has function of tumor suppression. In cytosol, histamine is metabolized by histamine-N- methyltransferase (HNMT). The concentration of histamine and its products have association with glioma. However, there is no direct evidence to investigate the roles of HNMT in growth and pathogenesis of glioma. Our data demonstrate that either specisement of GBM or glioma cell lines express more HNMT than normal brain tissue. More amount of HNMT suggests less tumorspressive histamine. On the other hands, we also find that epidermal growth factor (EGF), a glioma inducing factor, increases the expression of HNMT and changes the location of HNMT in glioma cell line. Collectively, our data provide hint that HNMT is another noval theraptic target for deadly glioma.

中文摘要 (Abstract in Chinese) 1
英文摘要 (Abstract in Chinese) 3
目錄 (Contents) 5
第一章 緒論 (Introduction) 8
一、 神經膠質瘤 9
二、 HNMT 13
第二章 實驗材料與方法 (Materials and Methods) 16
一、 藥品試劑 17
二、 常用溶液 21
三、 常用儀器 23
四、 實驗方法 25
A. Brain tumor primary cell生長media製備 25
B. 配製 Collagenase mix 25
C. Collagenase digestion 26
D. 細胞繼代培養方法 27
E. 凍細胞流程 27
F. 解凍細胞方式 28
G. 反轉錄-聚合酶連鎖反應 RT-PCR 29
H. Real time PCR流程 31
I. 西方墨點法 33
J. 免疫組織染色 35
K. 共軛焦顯微鏡成像 35
第三章 實驗結果 (Results) 37
一、各種和癌症有關的標的基因和六種神經膠質瘤細胞株的關
係 38
二、病人神經膠質瘤檢體和正常組織中各種標的基因mRNA表
現量的較 40
三、病人神經膠質瘤檢體和正常組織中ANXA6、DCL1、HNMT、
GPX3及TAGLN 之mRNA表現量的比較 40
四、病人神經膠質瘤檢體和正常組織中HNMT之mRNA表現量
的較 41
五、比較病人所取得的檢體切片及正常腦組織切片中HNMT蛋白
質表現形 43
六、比較不同神經膠質瘤細胞株及正常腦組織HNMT的mRNA
及蛋白質表現形 43
七、比較不同經膠質細胞株在EGF處理下HNMT的蛋白質表現
情形 44
第四章 討論 (Discussion) 45
第五章 圖表 (Figures) 49
圖一 50
圖二 52
圖三 53
圖四 55
圖五 56
圖六 58
圖七 66
圖八 68
圖九 70
圖十 71
第六章 參考文獻 (Reference) 72
附錄 (Appendices) 77
一、細胞株之基本資料 78
二、臨床檢體之基本資料 79
三、臨床病人基本資料 80

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