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研究生:胡淑寶
研究生(外文):Shu-Bauh Hu
論文名稱:干擾素-α對肝癌細胞株之血管內皮生長因子與E-cadherin蛋白質表現所造成的影響
論文名稱(外文):Effects of Interferon-α on Vascular Endothelial Growth Factor and E-cadherin Expression in Liver Cancer Cell Lines
指導教授:柯成國柯成國引用關係
指導教授(外文):Chen-Guo Ker
學位類別:碩士
校院名稱:高雄醫學大學
系所名稱:藥學研究所碩士在職專班
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:53
中文關鍵詞:E-cadherinVEGF遠端轉移作用血管新生作用干擾素-α侵襲作用
外文關鍵詞:InvasionAngiogenesisInterferon-αMetastasisVEGFE-cadherin
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目的: VEGF 與E-cadherin,分別被視之為血管新生作用與遠端轉移作用之觀察指標。由VEGF 與E-cadherin 表現量的增加或減少,我們期待找到腫瘤治療中相關的判定預後的指標。本研究嘗試評估干擾素-α於試管內對人類肝癌細胞株之腫瘤發生作用的影響效應。

材料與方法:採用兩列分化程度良好的人類肝癌細胞株,Hep G2, Hep 3B,一列分化程度差的人類肝癌細胞株與SK-Hep I。細胞株經以干擾素-α處置培養,評估干擾素-α對其腫瘤遠端轉移能力的影響。我們用不同濃度的干擾素-α(5000 unit/m; 1000 unit/ml),以不同時間(3;6天)處置Hep G與SK-Hep I。隨後,以ELISA test分析測試培養基中VEGF表現量。Hep 3B細胞株用對照組,以及分別含干擾素-α1000, 5000, 20000 unit/ml之新鮮培養基培養3天。之後,用西方墨點法與Immunocytochemistry(免疫細胞化學)確認細胞膜上E-cadherin 表現量的變化。除上述檢測外,這三株細胞株均經細胞計數與MTT測試以觀察其經干擾素-α處置後的存活率。

結果:在血管新生作用的觀察,在A組(5000 unit/m; 3天)與B組(1000 unit/ml, 6天),兩組均可見干擾素-α調降VEGF蛋白質表現量。A組:VEGF蛋白質表現量為,Hep G2 (無 IFN-α�恁G124.4,經IFN-α�捖B置:76.8),SK-Hep-I (無IFN-α�恁G44.5,經IFN-α�捖B置:30.0). B組,VEGF蛋白質表現量為,Hep G2 (無IFN-α:225.5,經IFN-α�捖B置:151.4),SK-Hep-I (無IFN-α�恁G74.1,經IFN-α�捖B置:53.4)。在細胞侵襲作用方面,西方墨點法與Immunocytochemistry(免疫細胞化學) 顯示干擾素-α調升E-cadherin 表現量。細胞株經干擾素-α處置後均呈現細胞計數與MTT 測試之細胞存活率下降,顯示干擾素-α對肝癌細胞株有抗細胞增殖作用。

結論:干擾素-α的抗腫瘤活性,至少部分來自於調降VEGF蛋白質表現量之抗血管新生作用,以及調升細胞膜上E-cadherin蛋白質表現量之抗細胞侵襲作用,細胞存活率下降之抗細胞增殖作用。藉本研究結果將來可再探討干擾素-α在治療計劃中適當的投與量,或在化學預防方面扮演起預防癌症復發的角色。
Purpose: Vascular endothelial growth factor(VEGF) and E-cadherin
have been examined and taken for indicators of angiogenesis and
metastasis respectively. In our study, we evaluated the in vitro effects of
IFN-α treatment on the tumorigenicity of liver cancer cell lines. From the
changes of the expression of VEGF and E-cadherin, we expected to
evaluate the prognostic markers.
Materials and Methods: Two well differentiated (Hep G2, Hep 3B)
and one poor differentiated (SK-Hep I) human liver cancer lines were
used. Cells were incubated with Interferon-α to evaluate their metastatic
potential. Hep G2, SK-Hep-I cells were incubated with IFN-α for
different concentrations (5000 unit/ml; 1000 unit/ml) and different
periods(3;6 days) of time. Then, the VEGF protein expression was
assayed by ELISA test in those culture media.
The Hep 3B cells were treated with fresh medium, that contained 1000,
5000, 20000 unit/ml INF-α. It were incubated for 3 days. After that,
E-cadherin expression changes was confirmed by Western blotting and
Immunocytochemistry. All three cell lines, treated with INF-α, their
survival rates were measured by cell counting and MTT test .
Results: In the angiogenesis study, both in group A(5000 unit/ml;
3days)and group B(1000 unit/ml, 6days), the VEGF protein expression
was down-regulated by IFN-α. In Group A, the VEGF protein expression
were, in Hep G2 (no IFN-α:124.4, with IFN-α: 76.8),and in SK-Hep-I (no
IFN-α: 44.5, with IFN-α: 30.0). In Group B, VEGF protein expression
were, in Hep G2 (no IFN-α: 225.5, with IFN-α:151.4) and SK-Hep-I (no
IFN-α: 74.1, with IFN-α: 53.4). In the cell invasion study, Western
blotting and Immunocytochemistry showed that E-cadherin expression
were up-regulated by the increasing concentration of IFN-α. The declined
cell number counts and survival rates of all three cell lines showed
antiproliferative activity of IFN-α.
Conclusion: IFN-α confers its antitumor activity, at least in part, by its
antiangiogenic activity, which results from decreased VEGF expression,
and by anti-invasive activity, which results from elevated E-cadherin
expression and antiproliferative activity on cancer cell lines. The results
may help us to design the treatment protocol and to define the
chemopreventive role of IFN-α.
Key words:Interferon-α,Angiogenesis,Invasion,Metastasis,VEGF,E-cadherin
目錄
中文摘要..……………………………………………………… 1
英文摘要……………………………………………………… …3

本文
壹、 緒論………………………………………………………. 5
貳、 材料與方法……………………………………………….17
參、 結果……………………………………………………….29
肆、 討論…………………………………………………….. 40
伍、 結論…………………………………………………….…46
陸、 參考文獻………………………………………………….47
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