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博碩士論文 etd-0725114-131202 詳細資訊
Title page for etd-0725114-131202
論文名稱
Title
探討ABT-751對肝細胞癌Huh7的自噬與凋亡之相互關係
Studies on the relationship between autophagy and apoptosis in hepatocellular carcinoma-derived cells Huh7 by treatment with ABT-751
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
53
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2014-07-28
繳交日期
Date of Submission
2014-08-25
關鍵字
Keywords
自噬作用、細胞凋亡、微管、肝癌、ABT-751
Hepatocellular carcinoma, Autophagy, Apoptosis, Microtubule, ABT-751
統計
Statistics
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中文摘要
本論文研究的目的為探討ABT-751對肝癌細胞中的細胞凋亡及自噬作用之影響。肝癌(Hepatocellular carcinoma, HCC)在世界上是最常見的惡性腫瘤之一,其在台灣近年來是造成癌症死因的第二位。目前用於抗癌的藥物不外乎針對腫瘤血管新生的抑制及微管合成的抑制。ABT-751就是利用抑制微管合成的方式使細胞停留在G2/M期,進一步誘導細胞凋亡產生,故本研究的乃欲探討ABT-751此種藥物對肝細胞癌Huh7的自噬與凋亡之交互關係。
本研究利用MTT 試驗得知,ABT-751在48 小時的IC50為1.5 μM,藉由Cyto-ID、Acridine orange staining及Immunoblotting分析,在ABT-751的處理之下,會誘導Huh7細胞啟動自噬作用此種機制並促使細胞的存活。為了得知是否抑制自噬作用能增加Huh7細胞的凋亡,利用自噬抑制劑3-Methyladenine (3-MA)及bafilomycin A1 (BafA1) 的處理可發現抑制自噬作用確實會增加Huh7細胞的凋亡。由以上試驗結果得知,ABT-751除了抑制微管的合成,還會誘導自噬作用的機制,若能抑制ABT-751所誘導的自噬作用,其將會提升對抗癌細胞的效果。
Abstract
The objective was to study the effects of ABT-751 on apoptosis and autophagy in hepatocellular carcinoma-derived cells. Hepatocellular carcinoma (HCC) is a common malignant tumor in the world. In recent years, HCC was the second leading cause of cancer death in Taiwan. Recently, the effects of anti-cancer drugs usually are mediated by the inhibition of tumor angiogenesis and microtubule synthesis. ABT-751 arrest cells in the G2/M phase due to its inhibition of microtubule synthesis and induction of cell apoptosis. The objective of thesis is to study the relationship between autophagy and apoptosis in hepatocellular carcinoma-derived cells Huh7 by treatment with ABT-751.
In this study, the cytotoxicity of ABT-751 to Huh7 cells was measured by MTT assay at 570 nm. The results indicated the IC50 of Huh7 treated with ABT-751 for 48 h was 1.5 µM. The assay of Cyto-ID, acridine orange staining and immunoblotting were used to determine the ABT-751-induced autophagy in Huh7 cells and the autophagy induced by ABT-751 has a role in cell survival. To investigate whether inhibition of autophagy could enhance apoptosis in Huh7 cells after the treatment of ABT-751. The results showed the inhibition of autophagy could enhance the effect of inducing Huh7 cells death due to the combinations of ABT-751 with the autophagy inhibitors 3-methylamphetamine (3-MA) or bafilomycin A1 (BafA1). Above results indicated that ABT-751 could inhibit microtubule polymerization and lead to autophagy, which promoted survival of cancer cells. Consequently, combinations of ABT-751 with the autophagy inhibitor, which will promote the effect of ABT-751 to induce cytotoxin.
目次 Table of Contents
Approval page i
Acknowledgements ii
Abstract
Chinese iii
English iv
Contents vi
List of figures vii
List of tables viii
Introduction 1
Materials and Methods 12
Results 20
Figures 27
Discussion 33
References 41
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