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Functional and Structural Analysis of Cell Death by Chemotherapeutic Agents in Human Kidney Cells
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 최상돈 | - |
| dc.contributor.author | 신현준 | - |
| dc.date.accessioned | 2018-11-08T08:22:52Z | - |
| dc.date.available | 2018-11-08T08:22:52Z | - |
| dc.date.issued | 2017-08 | - |
| dc.identifier.other | 25649 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/13388 | - |
| dc.description | 학위논문(박사)--아주대학교 일반대학원 :분자과학기술학과,2017. 8 | - |
| dc.description.tableofcontents | INTRODUCTION 1 Chapter 1. Doxorubicin induces necrosis through PARP1 dependently mitochondrial hyperactivation and an ATP production pathway, without p53 dependence 4 1.1 Abstract 4 1.2 Introduction 5 1.3 Results 7 DOX induced the expression of PARP1, p53, and DNA damage response (DDR) proteins 7 Mitochondrial hyperactivation and cATP production raised during DOX-induced cell swelling and necrosis 7 DOX induced DNA damage, cell cycle arrest, and oxidative stress independent of p53 8 PARP1 induced mitochondrial hyperactivation/cATP production and cROS generation, but did not alter mROS and NO generation 9 Generation of cATP triggered DNA damage and necrosis, but did not induce cROS or mROS generation 9 PARP1 inhibition reduced necrosis and plasma membrane rupture 10 Knockdown of PARP1 suppressed DOX-induced cATP production, cROS generation, and necrosis 10 1.4 Discussion 12 1.5 Methods 16 1.6 Figure and Figure legends 21 Chapter 2. Mechanisms of ROS and ERK during etoposide-induced cytotoxicity in human kidney cells 36 2.1 Abstract 36 2.2 Introduction 37 2.3 Results 39 ETO triggered activation of MAPKs, nucleus swelling and DNA damage 39 ETO triggered mitochondrial biogenesis, cytotoxicity and DNA damage by ROS generation 39 ETO triggered cell death and DNA damage by activation of ERK 40 ETO triggered NE ruptures through caspases activation by ERK 40 2.4 Discussion 42 2.5 Methods 44 2.6 Figure and Figure legends 50 DISCUSSION 57 REFERENCE 59 Chapter 3. APPENDIX 66 1. Paper 1: Doxorubicin Induces Cytotoxicity through Upregulation of pERK–Dependent ATF3 66 2. Paper 2: Etoposide Induces Necrosis through p53-mediated Antiapoptosis in Human Kidney Proximal Tubule Cells 77 3. Paper 3: Activating transcription factor 3 represses inflammatory responses by binding to the p65 subunit of NF-κB 94 4. Paper 4: Structural and functional analysis of cell adhesion and nuclear envelope nano-topography in cell death 104 5. Paper 5: Doxorubicin-induced necrosis is mediated by poly-(ADP-ribose) polymerase 1 (PARP1) but is independent of p53 119 6. Paper 6: Etoposide induced cytotoxicity mediated by ROS and ERK in human kidney proximal tubule cells 137 7. Paper 7: TLR4/MD2 specific peptides stalled in vivo LPS-induced immune Exacerbation 151 Chapter 4. 국문요약 164 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | Functional and Structural Analysis of Cell Death by Chemotherapeutic Agents in Human Kidney Cells | - |
| dc.title.alternative | Functional and Structural Analysis of Cell Death by Chemotherapeutic Agents in Human Kidney Cells | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | HYEON-JUN SHIN | - |
| dc.contributor.department | 일반대학원 분자과학기술학과 | - |
| dc.date.awarded | 2017. 8 | - |
| dc.description.degree | Doctoral | - |
| dc.identifier.localId | 788439 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000025649 | - |
| dc.subject.keyword | Cell Death | - |
| dc.description.alternativeAbstract | Cell death was generally classified into two forms: apoptosis, also known as the regulated cell death, and necrosis known as the unregulated cell death generally induced by chemotherapeutic drug and other stimulators. Apoptosis is most distinguished from morphologically necrosis by its major morphological features. For instance, apoptotic cells show cell shrinkage, apoptotic bodies and pyknosis meanwhile, necrotic cells reveal nuclear and cell swelling as well as plasma membrane rupture. Additionally, recent study described that tumor necrosis factor (TNF) induced necrosis through RIP in caspase 8 deficient cells. These findings imply the possibility that necrosis can also be regulated and this mechanism of cell death it called programmed necrosis or necroptosis. Numerous chemotherapeutic drugs are being developed or used for the treatment of cancer or other diseases, but these drugs have the limitation of target selection or serious side-effects. To improve the drug screening or effect, it is important to understand the cell death mechanism. In this study, we established renal dysfunction mechanism using topoisomerase II target drugs including doxorubicin and etoposide. Doxorubicin triggered the poly-(ADP-ribose) polymerase 1 (PARP1)-dependent cell death by necrosis but did not seem to effect activation of p53. Furthermore, etoposide induced cytotoxicity through reactive oxygen species (ROS) and extracellular signal regulated kinase (ERK) in human kidney proximal tubule (HK-2) cells. The first part of this study presented that the treatment of the HK-2 cells by doxorubicin induced DNA damage, PARP1, p53 activation and mitochondrial hyperactivation including increased mitochondrial respiration and outer membrane potential. Moreover, doxorubicin triggered the generation of nitric oxide (NO), cytosolic ROS (cROS), mitochondrial ROS (mROS), production of cytosolic adenosine triphosphate (cATP) and morphological changes including cell swelling and plasma membrane rupture. However, PARP1-inhibited cells reduced the doxorubicin-induced necrosis, mitochondrial hyperactivation, ROS generation, ATP production and DNA damage. In other hand, p53 deficient cells did not present a reduced t doxorubicin-induced DNA damage, cell cycle arrest and the production of oxidative stress factors such as cROS, mROS and NO. These results presented that doxorubicin-induced necrosis mediated by PARP1, but was p53 independent. The second part of this study presented that the treatment of HK-2 cells by etoposide triggered DNA damage, PARP1 and mitogen-activated protein kinases (MAPKs) activation including extracellular signal-regulated kinase-1 (ERK), p38 and c-Jun N-terminal kinase (JNK). Additionally, etoposide induced mitochondrial biogenesis, generation of ROS and production of cATP. The treatment of cells by N-acetylcysteine (NAC) and ROS scavenger, resulted in decreased etoposide-induced cell death by necrosis, a limited DNA damage and mitochondrial biogenesis, but did affect the activation of ERK, caspase 3 and apoptotic cells. In other hand, ERK-deficient cells reduced etopside-induced nuclear envelope ruptures, DNA damage, caspase 3/7 activity and cytotoxicity. Taken together, these findings showed that etoposide triggered the activation of ERK and the generation of ROS, which regulate differently cell death mechanism in HK-2 cells. ERK activation leads to caspase 3/7 activation which in return induces the nuclear envelope ruptures, and triggers cell death by apoptosis. Whereas, ROS generates mitochondrial biogenesis and the production of cATP, which lately induces cell death by necrosis, rather than apoptosis. | - |
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