出版年:

2000

研究生:

蔡靜姍

研究生(英文姓名):

Tsai, Ching-Shan

論文名稱:

以腺病毒攜帶抑制血管新生因子行小鼠膀胱癌基因治療

英文論文名稱:

Bladder cancer gene therapy using adenoviral vectors encoding angiogenic inhibitors

指導教授:

蕭璦莉

指導教授(英文姓名):

Shiau, Ai-Li

學位類別:

碩士

校院名稱:

國立成功大學　

系所名稱:

微生物暨免疫學研究所

學號:

s46871102

學年度:

88

語文別:

中文

論文頁數:

0

關鍵詞:

基因治療 ; 膀胱癌 ; 抑制血管新生

英文關鍵詞:

gene therapy ; bladder cancer ; anti-angiogenesis

被引用次數:

1

[ 摘要 ]

腫瘤的生長與轉移均需要靠血管新生的過程才能完成，因此抑制腫瘤的血管新生已經成為治療癌症的策略之一。Angiostatin （AST）與thrombospondin-1 (TSP-1)分別是plasminogen中的kringle片段與extracellular matrix中的主要成分，在過去的研究中指出此兩種蛋白質具有抑制新血管生成的能力。因為膀胱癌腫瘤比起正常的膀胱上皮細胞有顯著血管新生的現象，因此抑制血管新生的因子，如：angiostatin與TSP-1可能可以成為有效治療膀胱癌的物質。本研究的目的是在分析帶有angiostatin與TSP-1兩段cDNA的腺病毒載體對膀胱癌治療的效果。將小鼠的angiostatin與TSP-1 cDNA分別接到E1基因缺陷的腺病毒載體中，將其分別送入會提供E1基因的293細胞中，產生重組的腺病毒，將之大量繁殖並定其效價；並利用聚合連鎖反應與西方點墨法證明AST基因與蛋白質均在重組腺病毒中存在與表現。這兩種重組腺病毒可以抑制內皮細胞增生與遊走(migration)的能力；而在體內試驗中，它們會阻斷雞胚胎絨毛膜(chorioallantoic membrane)上的血管新生。應用在小鼠膀胱癌的基因治療上，結果顯示：帶有AST基因的重組腺病毒具有明顯抑制C3H/HeN小鼠膀胱癌腫瘤的生長，但帶有TSP-1基因的重組腺病毒則無抑制腫瘤生長的效果。總結來說，利用帶有血管新生抑制因子，如AST的重組腺病毒治療膀胱癌腫瘤為一個可行的方法。

[ 英文摘要 ]

Tumor growth and metastasis are angiogenesis-dependent. Anti-angiogenesis has been exploited as one of the strategies in cancer gene therapy. Angiostatin, the kringle region of plasminogen, and thrombospondin-1, an extracellular matrix glycoprotein, have been demonstrated as potent inhibitors of neovascularization. Since bladder tumors are characterized by markedly increased angiogenesis when compared to the normal urothelium from which they are derived, angiogenic inhibitors, such as angiostatin and TSP-1, may be promising agents for the treatment of bladder cancer. The aim of this study is to analyze the effectiveness of angiostatin and TSP-1 gene therapy, delivered by E1-deleted adenoviral vectors, for the treatment of murine bladder cancer. Mouse angiostatin kringle1-5 [AST (k1-5)] and TSP-1 cDNAs have been constructed into Ad5L adenoviral vector and the corresponding recombinant adenoviruses were generated in the packaging 293 cell line providing E1 in trans. After virus amplification, the virus titers were determined. The angiostatin cDNA and protein were detected in the viral particles by PCR and western blot analysis, respectively. The proliferation and migration of endothelial cells towards angiogenic factors were inhibited by the viruses. Furthermore, in vivo, they blocked neovascularization in chick embryo chorioallantoic membrane (CAM). These two replication-defective adenoviruses were used for bladder cancer therapy in the mouse bladder tumor model. C3H/HeN mice bearing bladder tumor were treated with Ad/AST or Ad/TSP-1 viruses. The results showed that Ad/AST viruses suppressed the tumor growth, whereas Ad/TSP-1 had no effect. In conclusion, adenoviruses expressing angiogenic inhibitors, such as angiostatin, may be effective approaches in the treatment of bladder cancer.