인간 대장암에서 AMPK/ACC 신호전달 경로를 통한 뷰티레이트에 의한 내성유도에 관한 연구
Butyrate-induced resistance through AMPK/ACC signaling pathway in human colon cancers
- 주제(키워드) butyrate , resistance , colon cancer , AMPK , AICAR
- 발행기관 아주대학교
- 지도교수 김소희
- 발행년도 2020
- 학위수여년월 2020. 2
- 학위명 석사
- 학과 및 전공 일반대학원 약학과
- 실제URI http://www.dcollection.net/handler/ajou/000000029692
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Butyrate, a short-chain fatty acid produced by the colonic bacterial fermentation, inhibits cell growth in human colon cancer cells through the inhibition of histone deacetylases. However, chronic exposure to butyrate induces the resistance to butyrate in human colon cancer cells. The mechanism for the acquisition of resistance has not yet been fully understood. Butyrate-resistant (BR) cancer cells were developed by gradually increasing the butyrate concentration up to 1.6 mM for approximately 3 months in HCT116, HT29 and SW480 human colon cancer cells. The butyrate concentrations that inhibited cell growth by 50% (IC50) were increased by 11.8, 13.2 and 6.10-fold in HCT116, HT29 and SW480 BR cells, respectively, then their parental (PT) cells. Butyrate-resistance induced the autophagy through higher expression of Beclin-1 and LC3B. The expression of AMP-activated protein kinase (AMPK) was down-regulated in BR cells along with the activation of AKT and mTOR compared to respective PT cells. Decreased AMPK expression resulted in the expression of enzymes involved in fatty acid synthesis, such as acetyl-CoA carboxylase (ACC) and ATP citrate lyase (ACLY) in BR cells. Activation of AMPK by treating 5-Aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR, 1 mM) suppressed cell proliferation by inhibiting. Taken together, chronic butyrate exposure increased resistance to butyrate in the human colon cancer through down-regulation of AMPK/ACC signaling pathway, suggesting that AMPK could be a therapeutic target for BR colon cancers.
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ABSTRACT i
TABLE OF CONTENTS ii
LIST OF FIGURES iv
LIST OF TABLES v
ABBREVIATIONS vi
I. INTRODUCTION 1
II. MATERIALS AND METHODS 2
A. Reagents 2
B. Cell culture and establishment of butyrate-resistance cells 2
C. Cell proliferation assay 2
D. Flow cytometry 3
E. Immunofluorescence analysis 3
F. RNA extraction and qRT-PCR 3
G. Immunoblot analysis 4
H. Statistical analysis 4
III. RESULTS 6
A. Effects of butyrate on the cell proliferation of colon cancer 6
B. Effects of butyrate-resistance on drug efflux pumps expression 9
C. Effects of butyrate-resistance on the cell cycle and apoptosis 11
D. Induction of autophagy by butyrate-resistance 14
E. Effects of butyrate-resistance on AMPK/ACC and AKT/mTOR pathway 16
F. Effects of butyrate-resistance on expression of fatty acid synthesis enzymes 18
G. Activation of AMPK/ACC pathway in BR cells 20
IV. DISCUSSION 23
V. CONCLUSION 24
REFERENCES 26
KOREAN ABSTRACT 29
