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The study for the functional role of MTH1 against oxidative stress

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Author(s)
송승희
Issued Date
2006
Abstract
정상 세포의 호흡과정중에 발생되는Oxygen radicals은 genomic과 mitochondrial DNA 에서의 mutations을 일으킬수 있다. Human MTH1은 8-oxo-dGTP나 2-hydroxy-dATP와 같은 산화된 purine nucleoside triphosphates를 가수분해하여, 복제 동안 이들 산화 nucleotide의 삽입 착오를 방지한다. MTH1에 대해서 선택적인 small interfering RNAs (siRNAs) 가 설계, 합성, 사용되었고, DNA 손상에 대한 세포의 생존능력 전환 및 H2O2 에 대한 세포의 저항성에 있어서의 효율성이 연구되었다. 인간 섬유아세포에서 MTH1 발현 억제는 H2O2 에 대항하는 세포 생존능력을 감소시킨다. 게다가 MTH1 발현 억제는 H2O2 농도에 따라서 세포 죽음을 증가시킨다. 또한 MTH1 결핍세포에서 H2O2 투여 후 caspase 와 PARP의 절단이 증가함을 관찰하였다. 이상의 연구결과 산화성 유전자 손상 복구 효소인 MTH1은 산화성 스트레스에 의한 세포 사멸사 억제에도 관여할 것으로 사료된다.|Oxygen radicals generated through normal cellular respiration process can cause mutations in genomic and mitochondrial DNA. Human MTH1 hydrolyzes oxidized purine nucleoside triphosphates, such as 8-oxo-dGTP and 2-hydroxy-dATP, to monophosphates, thereby preventing the misincorporation of these oxidized nucleotides during replication. We designed, synthesized, and utilized small interfering RNAs (siRNAs) that were selective for MTH1 and investigated their effectiveness in altering the viability of the cells against DNA damage as well as the resistance of the cells to H2O2. The suppression of MTH1 expression in the human fibroblast cells lead to a decrease in the cell viability against H2O2. In addition, the cell death was increased concentration-dependent in the suppression of MTH1 expression. Moreover, MTH1 knockout cells have shown that caspase and PARP are fragmented after exposure of H2O22 but, not control cells. These results suggest that MTH1, which is well known to repair the oxidative bases, also has an antiapoptotic function against oxidative stress.
Alternative Title
산화성 손상에 대한 MTH1 역할에 관한 연구
Alternative Author(s)
Song, Seung hee
Affiliation
조선대학교 대학원
Department
일반대학원 생물신소재학과
Advisor
유호진
Awarded Date
2006-02
Table Of Contents
KOREAN ABSTRACT = 1
ABSTRACT = 2
I. INTRODUCTION = 3
II. MATERIALS AND METHODS = 5
Cell culture = 5
Preparation of the constructs and clones = 5
siRNA design, synthesis, and expression vector transfected cell clones = 5
Western blotting = 7
Semiquantitative Reverse Transcriptase Polymerase Chain Reaction = 7
Flow cytometry = 8
Microarray analysis = 9
Cytotoxicity assays = 10
III. RESULTS = 12
siRNA mediated down regulation of MTH1 = 12
Silencing of MTH1 expression via siRNA leads to a decrease in the cell viability capacity in response to H2O2 = 13
Silencing of MTH1 expression via siRNA leas to a increase in the cell death in response to H2O2 = 14
Cell death by the suppression of MTH1 gene expression related the apoptosis = 14
Analysis of gene expression by biological function using microarray and RT PCR = 15
IV. DISCUSSION = 27
V. REFERENCES = 33
Degree
Master
Publisher
조선대학교 대학원
Citation
송승희. (2006). The study for the functional role of MTH1 against oxidative stress.
Type
Dissertation
URI
https://oak.chosun.ac.kr/handle/2020.oak/6085
http://chosun.dcollection.net/common/orgView/200000232746
Appears in Collections:
General Graduate School > 3. Theses(Master)
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