DNA damaging agent에 의한 유방암 세포 사멸에서 RIP3의 역할
Role of RIP3 in DNA damaging agent-induced breast cancer cell death
- 주제(키워드) RIP3 , MLKL , Programmed necrosis , Hypomethylating agents , Chemotherapy
- 발행기관 아주대학교
- 지도교수 김유선
- 발행년도 2015
- 학위수여년월 2015. 8
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000020500
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Receptor-interacting protein kinase 3 (RIP3 or RIPK3) is a central player in “programmed” or “regulated” necrotic cell death pathway. Here we show that programmed necrosis is activated in response to chemotherapeutic agents and contributes to chemotherapy-induced breast cancer cell death. However, we also show that RIP3 expression is silenced in basal like breast cancer cell lines due to DNA methylation near its transcription start site. Due to this silencing mechanism, loss of RIP3 expression in these cells leads to resistance to DNA damaging-agents. Hypomethylating agents restore RIP3 expression and promote sensitivity not only to death receptor ligands, but also to a surprising diversity of standard chemotherapeutic agents in a RIP3-specific manner. Our data indicates that RIP3-dependent breast cancer cell death is activated in response to DNA damaging-agents. This suggests that RIP3 plays a greater role in response to DNA damaging-agents than has been previously appreciated, we propose that hypomethylating agents, in combination with standard chemotherapy, may be useful in treating breast cancers that lack RIP3 expression.
more목차
I. INTRODUCTION 1
II. MATERIALS AND METHODS 4
A. Experimental reagents 4
B. Cell Culture 4
C. Lentiviral shRNA experiments 4
D. Reverse Transcription-PCR 5
E. Cell viability assay 5
F. Western blot analysis 5
G. Immunoprecipitation 6
H. Pyrosequencing of Bisulfate converted genomic DNA 6
I. Colony formation assay 6
J. Invasion assay 7
III. RESULTS 8
A. RIP3 expression is often silenced in breast cancer cells due to genomic methylation near its transcriptional start site 8
B. Expression of RIP3 is essential for TNF-induced necrotic cell death in cancer cells 13
C. RIP3 expression sensitizes to DNA-damaging agents in breast cancer cells 18
D. DNA-damaging agents activate RIP3-dependent MLKL phosphorylation 24
E. RIP3 acts as tumor suppressor 27
IV. DISCUSSION 31
REFERENCES 33
국문요약 39
