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博碩士論文 etd-0118116-092731 詳細資訊
Title page for etd-0118116-092731
論文名稱
Title
探討ABT-751在肝癌細胞株Hep-3B經由ROS造成細胞凋亡及自噬作用
Studies on ABT-751-induced Apoptosis and Autophagy via Reactive Oxygen Species in the Hepatocellular Carcinoma-derived Hep-3B cells
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
70
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
2016-01-27
繳交日期
Date of Submission
2016-02-18
關鍵字
Keywords
ABT-751、細胞凋亡、DNA損傷、Caspase-dependent pathway、活性氧化物、⾃噬作⽤
Caspase-dependent pathway, Autophagy, Apoptosis, DNA damage, ROS, ABT-751
統計
Statistics
本論文已被瀏覽 5686 次,被下載 5
The thesis/dissertation has been browsed 5686 times, has been downloaded 5 times.
中文摘要
肝細胞癌在癌症中是具極惡性的癌症。全身性化學治療在肝癌實質細胞中也是最困難的,抗有絲分裂藥物在目前的治療上仍積極研究中。在本研究中,我們顯示ABT-751在抗癌活性的影響及其是如何造成Hep-3B細胞死亡之機轉。ABT-751是一種抗有絲分裂的藥物它可以使Hep-3B細胞的細胞週期停滯、微小管的去聚合反應以及抑制細胞的非貼附性生長。本實驗也發現ABT-751提高細胞產生活性氧化物及降低粒線體膜電位。除此之外,ABT-751活化Caspase-dependent相關蛋白以及導致BAX和BAK轉位至粒線體膜上,進而影響粒線體膜的通透性使得cytochrome c釋出到細胞質中。我們也發現ABT-751抑制凋亡誘導因子的活化。在我們的結果中,N-acetylcysteine阻斷ABT-751誘導活性氧化物生成造成的Hep-3B細胞凋亡。值得注意的是在先前研究中ABT-751不會誘導Hep-3B細胞產生自噬作用,但是在我們的實驗發現將TP53 null的Hep-3B細胞中overexpression TP53後,ABT-751會誘導其產生自噬作用。總結以上的發現,ABT-751是藉由產生活性氧而誘導Hep-3B細胞凋亡。
Abstract
Hepatocellular carcinoma (HCC) is one of the most frequently lethal malignant tumors in the world. Systemic therapies in hepatocellular carcinoma of solid tumors, one of the most difficult, and the anti-mitotic chemotherapy drug in the treatment of liver cancer is still active study. In the study, we performed the effect of antitumor activity at ABT-751-treated cells and further elucidated the mechanism of ABT-751-induced cell death in Hep-3B cells. ABT-751 is an antimitotic drug, which in our results it was induced cell cycle arrest, inhibited microtubules polymerization and decline anchorage-independent growth in Hep-3B cells. We also observed ABT-751 up-regulated Reactive oxygen species (ROS) generation and down-regulated mitochondria membrane potential (MMP) in Hep-3B cells. In addition, ABT-751 cleaved caspase-dependent related proteins and caused BAX and BAK translocation to mitochondrial membrane, followed mitochondrial membrane permeability increasing and cytochrome c releasing. Further, we examined apoptosis-inducing factor (AIF) which is caspase-independent related protein is decreased in nucleus by analyzing ABT-751-treated cells. In our results, ABT-751 was decreased apoptosis by N-acetylcysteine (NAC) blocking ROS generation. It is worth noting that ABT-751 did not induce autophagy in previous studies in Hep-3B cells (TP53 null cell line). However, we were demonstrated ABT-751 inducing autophagy in Hep-3B cells with transfected pTP53-HaloTag. Above these results, ABT-751 is induced cell apoptosis though the generation of reactive oxygen species in Hep-3B cells.
目次 Table of Contents
Approval page i
Acknowledgement ii
Abstract iii
Chinese iii
English iv
List of figures vii
List of tables viii
Introduction 1
Materials and Methods 11
Results 27
Figures 33
Discussion 45
Supplementary Data 52
References 57
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