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研究生:蘇郁晴
研究生(外文):Yu-Ching Su
論文名稱:Doxorubicin引發tNOX表現量增加導致A549肺癌細胞移動情形增加
論文名稱(外文):Doxorubicin-mediated transient tNOX up-regulation leading to enhanced cell migration in A549 lung cancer cells
指導教授:闕斌如
口試委員:林芸薇莊秀美
口試日期:2011-07-02
學位類別:碩士
校院名稱:國立中興大學
系所名稱:生物醫學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:英文
論文頁數:31
中文關鍵詞:腫瘤相關NADH氧化細胞移動
外文關鍵詞:tNOXdoxorubicincell migration
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tNOX (tumor-associated NADH oxidase)是屬於一種與細胞生長有關的膜外-NADH氧化

tNOX (tumor-associated NADH oxidase) belongs to an ECTO-NOX family of growth-related NADH oxidases. Since over-expressing tNOX leads to cells transforming, enhanced invasion ability and exhibited more responsive to anticancer drugs. On the other hand, HeLa cell growth and migration are reduced by tNOX-knockdown utilizing RNA interference technique, implying that tNOX is associated with transformed cells proliferation, migration and invasive ability. Doxorubicin (adriamycin), a long-time use in clinical cancer treatment, has been demonstrated to be a target of tNOX. Here, we confirmed that the expression of tNOX is inhibited by doxorubicin treatments at both protein and RNA levels. Interestingly, the expression of tNOX is transiently enhanced by short-time exposure of lower concentrations of doxorubicin in association with increased A549 cells migration and proliferation, suggesting that the level of tNOX expression is important for aggressive cancer cell phenotypes. To explore tNOX function more deeply, we found a serine504 mutation on tNOX exhibits enhanced tNOX function in cell proliferation, migration and different responses against doxorubicin treatment compared to wild type tNOX.

中文摘要........................................................................................................................I
Abstract........................................................................................................................II
Catalog........................................................................................................................III
Chapter 1 Introduction................................................................................................1
Part1 tNOX (tumor-associated NADH oxidase) ...........................................................1
Background.................................................................................................................1
tNOX function............................................................................................................3
Part2 Doxorubicin (adriamycin) ....................................................................................4
Chapter 2 Materials and methods..............................................................................6
1. Cell culture and plasmids...........................................................................................6
2. Cell growth and cell migration...................................................................................7
3. Cell-cycle analysis......................................................................................................7
4. Western blot analysis..................................................................................................8
5. Reverse Transcriptase polymerase chain reaction......................................................8
6. Apoptosis assay...........................................................................................................9
7. Boyden chamber assay...............................................................................................9
8. ROS measurement......................................................................................................9
9. Statistics...................................................................................................................10
Chapter 3 Results.......................................................................................................11
1. A549 cells proliferation were inhibited by doxorubicin treatment. ........................11
2. Doxorubicin-mediated growth inhibition is caused by cell cycle arrest, but not apoptosis. ...................................................................................................................11
3. Expression of tNOX was down-regulated during low concentration of doxorubicin treatment, but transient up-regulated by short-time treatment. .................................12
4. Cell migration of A549 cells were enhanced with low concentration. ...................12
5. Knockdown of tNOX inhibited cells migration. .....................................................13
6. A mutation at Ser504 on tNOX exhibits enhanced tNOX function in migration and proliferation. ..............................................................................................................13
7. Wild type and mutant tNOX exhibit different response to doxorubicin in cell. .....14
8. S504A-tNOX interferes with NAC to generate more ROS. ...................................14
Chapter 4 Figures ......................................................................................................15
Chapter 5 Discussion .................................................................................................24
Chapter 6 References ................................................................................................26


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