Increased chemoresistance by butyrate in human colon cancer cells
- 주제(키워드) Colon cancer , Butyrate , Chemoresistance , Apoptosis , HCT116 cells
- 발행기관 아주대학교
- 지도교수 김소희
- 발행년도 2016
- 학위수여년월 2016. 2
- 학위명 석사
- 학과 및 전공 일반대학원 약학
- 실제URI http://www.dcollection.net/handler/ajou/000000021798
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Butyrate is a short-chain fatty acid produced by intestinal microflora and it not only induces apoptosis, but also inhibits cell proliferation in cancer. Recently, it has been reported butyrate may cause resistance in colon cancer. So we investigated the effects of increased resistance to butyrate in HCT116 colon cancer cells. We established HCT116 cells resistant to butyrate (HCT116/BR) by treating HCT116 parental cells (HCT116/PT) with increasing the concentration of butyrate gradually up to 1.6 mM for three months. The concentrations inhibiting the cell growth by 50% (IC50) of butyrate were 0.508 and 5.50 mM in HCT116/PT and HCT116/BR. The values of IC50 after the treatment with paclitaxel, 5-fluorouracil (5-FU), doxorubicin or trichostatin A (TSA) were 2.42, 2.36, 4.31 and 11.3 fold higher, respectively, in HCT116/BR cells compared with HCT116/PT cells. The protein expression of drug efflux pumps such as P-glycoprotein (P-gp), breast cancer resistant protein (BCRP) and the multidrug resistance associated protein 1 (MRP1) were not changed between HCT116/PT and HCT116/BR cells. The expressions of anti-apoptotic Bcl-xL protein were increased while those of pro-apoptotic Bax and Bim proteins were reduced in HCT116/BR cells. There were no significant differences in cell motility and invasion. Our study suggests that when colon cancer cells are exposed to butyrate consistently, cancer cells can have resistance to butyrate which may play a role in acquisition of chemoresistance in colon cancer.
more목차
I. INTRODUCTION 1
II. MATERIAL AND METHODS 3
A. Chemicals 3
B. Establishment of butyrate-resistant cell line 3
C. Cell proliferation assay 4
D. Flow cytometry 4
E. Protein extraction and Immunoblot assay 5
F. Gelatin zymography 6
G. Wound healing assay 6
H. Statistical analysis 7
III. RESULTS 8
A. Establishment of butyrate-resistant HCT116 cell lines 8
B. Chemoresistance in HCT116/BR cells 10
C. The protein expression of drug efflux pumps 13
D. Cell cycle arrest induced by butyrate 15
E. Protein expression on cell cycle and apoptosis 17
F. Effect of butyrate on invasiveness 21
IV. DISCUSSION 24
V. CONCLUSION 27
REFERENCES 28
KOREAN ABSTRACT 33
