Full metadata record
DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | 강상원 | - |
dc.contributor.author | 丁炯玎 | - |
dc.creator | 丁炯玎 | - |
dc.date.accessioned | 2016-08-26T12:08:06Z | - |
dc.date.available | 2016-08-26T12:08:06Z | - |
dc.date.issued | 2004 | - |
dc.identifier.other | OAK-000000009775 | - |
dc.identifier.uri | https://dspace.ewha.ac.kr/handle/2015.oak/191134 | - |
dc.identifier.uri | http://dcollection.ewha.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000009775 | - |
dc.description.abstract | TNF-α induces apoptosis via the conversion of procaspases to mature caspases, concomitant with the production of intracellular H₂O₂. In present study, I seek to elucidate the mechanism by which ROS cross talk with TNF-induced apoptosis. To achieve this goal, we utilized HeLa cells stably expressing a dominant negative mutant of a cytosolic peroxidase, peroxiredoxin Ⅱ, which is known to be a H₂O₂ signal regulator. The expression of Prx Ⅱ mutant 1) increased H₂O₂ production ; 2) enhanced cytochrome c relrease to cytosol and caspase 3 activation ; 3) exacerbated apoptotic cell death in response to TNF-α ; 4) enhanced caspase 8 activation. JNK activation was not involved in this cell death. In order to confirm this, I used RNA interference to known-down Prx Ⅱ in HeLa cells. Prx Ⅱ siRNA transfection increased TNF-induced apoptotic death and caspase activation in HeLa cells. Together, this study suggests H₂O₂ regulated by Prx Ⅱ somewhat amplifies caspase activation in response to TNF-α.;TNF-α는 procaspse를 활성화된 caspase로 전환시키는 동시에 H2O2의 생성을 수반하여 apoptosis를 유발한다. 이 논문에서는 ROS가 어떠한 기전으로 TNF에 의해 유도되는 apoptosis와 연관되어 있는지에 대해 연구하였다. 이를 밝히기 위해 H₂O₂의 signal regulator인 cytosolic peroxiredoxin, peroxiredoxin Ⅱ 의 dominant negative mutant를 안정하게 발현시키는 Hela 세포를 사용하였다. 이 Prx Ⅱ mutant 를 발현하는 cell은 Prx Ⅱ WT cell과는 반대로 TNF-α의 자극에 대해 민감한 반응을 나타내었다. 이러한 현상은 apoptosis의 초기 단계에서 caspase8의 활성화를 증가시키기 때문이었고, 이는 apoptosis의 외인성 및 내인성 경로를 모두 경유하는 것임을 밝혔다 .그리고 이것은 JNK 의 활성화와는 무관하다는 것을 알게 되었다. 이들을 종합해 볼 때, 이 논문에서는 PrxⅡ에 의해 조절되는 H2O2가 TNF에 의한 caspase의 활성에 어느 정도 영향을 준다는 것을 보여주고 있다. | - |
dc.description.tableofcontents | 목차 ABSTRACT = 1 TABLES OF ONTENTS = 3 LIST OF FIGURES = 5 1.INTRODUCTION = 6 2.MATERIALS AND METHODS = 11 2.1. Materials and Instruments = 11 2.2 Cell Culture = 12 2.3. Immunoblotting = 13 2.4. Apoptosis Assay = 14 2.5. Caspase activity assay = 14 2.6. Depletion of Prx II by RNAi = 15 2.7. Subcellular Fractionation = 16 2.8. Measurement of ROS = 16 2.9. Data Analysis = 16 3. RESULTS = 18 3.1.TNFa induces ROS generation in Hela cells = 18 3.2. Active site cysteine of Peroxiredoxin II is slightly inactivated after cell death = 21 3.3. Effects of expression of Prx II (WT) or Prx II (DN) on ROS generation and TNFa-induced cell death = 25 3.4. Apoptosis was inhibited not by JNK inhibitor but by caspase Inhibitor = 33 3.5. Intrinsic apoptosis pathway participates in ROS-modulated apoptosis = 37 3.6. Enhanced activation of initiator caspase in DN10 cells = 42 3.7. Effects of Prx II depletion by siRNA on apoptosis = 47 4. Discussion = 51 5. References = 55 6. 논문개요 (한글) = 64 | - |
dc.format | application/pdf | - |
dc.format.extent | 725700 bytes | - |
dc.language | eng | - |
dc.publisher | 이화여자대학교 대학원 | - |
dc.title | Regulatory role for Peroxiredoxin II in TNF-α-Induced Apoptosis | - |
dc.type | Master's Thesis | - |
dc.format.page | 65 p. | - |
dc.identifier.thesisdegree | Master | - |
dc.identifier.major | 대학원 분자생명과학부 | - |
dc.date.awarded | 2005. 2 | - |