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系統識別號 U0026-0407201216375600
論文名稱(中文) 探討順鉑或紫杉醇合併蟲草素對MA-10老鼠萊氏腫瘤細胞進行細胞凋亡之影響
論文名稱(英文) The Apoptotic Effect of Cisplatin and Taxol Combined with Cordycepin on MA-10 mouse Leydig Tumor Cells
校院名稱 成功大學
系所名稱(中) 細胞生物及解剖學研究所
系所名稱(英) Institute of Cell Biology and Anatomy
學年度 100
學期 2
出版年 101
研究生(中文) 陳姵蓉
研究生(英文) Pei-Jung Chen
學號 T96981063
學位類別 碩士
語文別 英文
論文頁數 70頁
口試委員 指導教授-黃步敏
口試委員-司君一
口試委員-江美治
中文關鍵字 蟲草素  順鉑  太平洋紫杉醇  加成性  合併處理  細胞凋亡 
英文關鍵字 Cordycepin  Cisplatin  Taxol  Drug combination  Additive effect  Apotosis 
學科別分類
中文摘要 萊氏細胞癌形成的主要原因目前還不是很清楚。近年來,化療不再限制於單一藥物的使用,很多研究都證實不同的抗癌藥物合併的確可以達到良好的效果。而且病人的反應也都相當良好。除此之外,抗癌藥物在細胞凋亡的影響上,除了會有形態上的變化,也會影響細胞週期,導致細胞週期的停滯。根據我們實驗室之前的研究顯示,蟲草素 (3'-脫氧腺苷)會藉由活化硫胱胺酸蛋白脢-3,-8,-9,進一步造成MA-10小鼠萊氏細胞癌的凋亡。所以,我們想探討順鉑、太平洋紫杉醇和蟲草素三種藥物經由不同組合的處理是否可在MA-10小鼠萊氏癌細胞上觀察到加成性的療效。根據實驗結果顯示,在順鉑、太平洋紫杉醇和蟲草素三種藥物經由不同濃度組合的處理下,MA-10小鼠萊氏癌細胞在形態上可觀察到細胞凋亡的情形,例如:細胞膜出泡。而在細胞存活試驗中 (MTT assay),細胞存活率在順鉑、太平洋紫杉醇和蟲草素三種藥物不同濃度合併使用下,效果比藥物單獨處理更加明顯。 流式細胞儀的檢測中,細胞在藥物的合併處理下,subG1時期的細胞累積量會比單獨處理更明顯。細胞機制的研究中,順鉑、太平洋紫杉醇和蟲草素三種藥物的不同組合可誘導硫胱胺酸蛋白脢-3, -8,-9活化以及PARP的裂解。同時在MAPKs的途徑研究中,JNK、ERK和 P38也都有被活化的情形。綜合以上結果得知,順鉑、太平洋紫杉醇和蟲草素會藉由活化內在/外在的硫胱胺酸蛋白脢以及MAPKs的途徑引發MA-10小鼠萊氏癌細胞的凋亡,並發現順鉑、太平洋紫杉醇和蟲草素藥物的不同組合使用可能具有潛在的加成性抗癌效果。
英文摘要 The causes of Leydig tumor cell still remain unknown. Recently, chemotherapies are not limited to single treatment anymore, and evidence suggests that different drug combinations are applied in patients with positive responses. Besides, the effects of anticancer drugs generally cause cell morphological changes and cell cycle arrest at different phases to induce cell apoptosis. Previous data in our laboratory have shown that cordycepin could induce MA-10 cell apoptosis by activating caspase-3, -8 and -9. Consequently, we investigated the possible additive and/or synergistic effect in apoptosis by cisplatin and/or taxol combined with cordycepin in MA-10 mouse Leydig tumor cell line. Results showed that MA-10 cells exhibited the apoptotic features, including cellular rounding-up and membrane blebbing, after treating with cordycepin, taxol, cisplatin, or cordycepin plus cisplatin and/or taxol for 24 hours. In MTT assay, drug alone or in combination treatments diminished MA-10 cell viability in a dose-dependent manner, and there were additive effect in various drug combinations, except for cisplatin plus taxol treatment. In flow cytometry assay, the treatments of cisplatin alone or cisplatin plus taxol induced cell arrest at G2/M phase without significant change in sub-G1 phase. However, other treatments displayed cell accumulation at sub-G1 phase. Moreover, the mechanism of apoptosis was detected by Western blotting. Data showed that caspase and MAPK and p53 signal pathways could be induced under the various treatments. Taken together, we found that cisplatin and/or taxol combined with cordycepin could induce an additive effect of apoptosis in MA-10 cells.
論文目次 Table of Contents
Abstract in Chinese……………………………………………….. i
Abstract……………………………………………………………. ii
Acknowledgements………………………………………………... iii
Table of Contents………………………………………………..... iv
List of Figures………………………………………………........... vi
Introduction……………………………………………….............. 1
Materials and Methods
Chemicals………………………………………………................... 8
Cells and cell culture……………………………………………….. 8
Morphology study………………………………………………….. 9
MTT cytotoxicity assay……………………………………………. 9
Apoptosis analysis by flow cytometry…………………………….. 10
Immunobloting analysis……………………………………………. 10
Statistical analysis………………………………………………….. 11
Results
Effects of taxol and/or cisplatin plus cordycepin on morphological change in MA-10 cells.……………………………………………...
12
Effects of taxol and/or cisplatin plus cordycepin on cell viability in MA-10 cells.………………………………………………………...
12
Effects of taxol and/or cisplatin plus cordycepin on cell cycle progression in MA-10 cells. ………………………………………..
13
Effect of taxol and/or cisplatin plus cordycepin on the protein expression of cleaved caspase-9 in MA-10 cells.…………………...
14
Effect of taxol and/or cisplatin plus cordycepin on the protein expression of cleaved caspase-8 in MA-10cells.……………………
15
Effects of taxol and/or cisplatin plus cordycepin on the protein expression of cleaved caspase-3 in MA-10 cells.…………………...
16
Effects of taxol and/or cisplatin plus cordycepin on the protein expression of cleaved PARP in MA-10cells.………………………..
17
Effects of taxol and/or cisplatin plus cordycepin on the protein expression of MAPKs in MA-10 cells..……………………………..
17
JNK inhibitor (SP600125) reversed cordycepin-induced phosphorylation of JNK in MA-10 cells.……………………………
19
JNK inhibitor (SP600125) reversed taxol and/or cisplatin plus cordycepin-induced phosphorylation of JNK in MA-10cells.………
20
ERK inhibitor (PD98059) reversed cordycepin-induced phosphorylation of ERK in MA-10 cells.…………………………...
20
ERK inhibitor (PD98059) reversed taxol and/or cisplatin plus cordycepin-induced phosphorylation of ERK in MA-10 cells.…......
20
P38 inhibitor (SB203580) reversed cordycepin-induced phosphorylation of p38 in MA-10 cells.…………………………….
21
P38 inhibitor (SB203580) reversed taxol and/or cisplatin plus cordycepin-induced phosphorylation of p38 in MA-10 cells.………
21
Effects of taxol and/or cisplatin plus cordycepin on the protein expression of p53 in MA-10 cells.…………………………………..
22
Discussion…………………………………………………………... 23
References………………………………………………………….. 28

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