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Functional and Structural Analysis of Cell Death by Chemotherapeutic Agents in Human Kidney Cells
- Alternative Title
- Functional and Structural Analysis of Cell Death by Chemotherapeutic Agents in Human Kidney Cells
- Author(s)
- 신현준
- Alternative Author(s)
- HYEON-JUN SHIN
- Advisor
- 최상돈
- Department
- 일반대학원 분자과학기술학과
- Publisher
- The Graduate School, Ajou University
- Publication Year
- 2017-08
- Language
- eng
- Keyword
- Cell Death
- Alternative Abstract
- 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.
- Fulltext
- Appears in Collections:
- Graduate School of Ajou University > Department of Molecular Science and Technology > 4. Theses(Ph.D)
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