Imidazole 항진균제 약물의 유방암에서의 PI3K-Akt기전을 통한 mTOR억제 효과에 따른 항암기전 연구
Imidazole Antifungal Drugs Induce Cell Death by Suppressing mTOR through PI3K-Akt Inhibition in Human Breast Cancer
- 주제(키워드) Azole계 항진균제 , 세포사멸 , cell cycle arrest , 오토파지 , PI3K-Akt-mTOR 기전
- 발행기관 아주대학교
- 지도교수 김소희
- 발행년도 2018
- 학위수여년월 2018. 2
- 학위명 석사
- 학과 및 전공 일반대학원 약학과
- 실제URI http://www.dcollection.net/handler/ajou/000000027580
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Breast cancer is a top woman cancer and an incidence rate increases every year. So it needs a perfect treatment. Azole drugs structurally have an imidazole [clotrimazole (CTZ) and ketoconazole (KCZ)] or a triazole [fluconazole (FCZ) and itraconazole (ICZ)]. Recently, azole antifungal drugs shown the anticancer efficacy in clinical trial. But, its antitumor mechanism are vague. Here, we evaluated the effects, as apoptosis, cell arrest, and autophagy and tumor growth using human breast cancer cells. Apoptosis assay shown imidazole induced cell death, pro-apoptosis maker, as cleaved PARP, cleaved caspase-3, Bax, increased and anti-apoptosis maker, as Bcl-2, decreased by CTZ and KCZ in both MCF-7 and MDA-MB-231 cells. And we also checked imidazole of azole drugs blocked the cell cycle in G0/G1 phage by down-controlling the protein levels of cyclin D1, CDK2, and CDK6 and increased p21 and p27 in breast cancer. Further experiments exhibited azole drugs also induced autophagy through the increased accumulation of LC 3B inside the cells by immunoblot analysis and immunofluorescence staining, increased beclin-1 and decreased p62/SQSTM1 protein. With azole decreased that though 3-MA. Azole drugs inhibited PI3K/Akt/mTOR signaling pathway. Tumor growth was also inhibited in nude mice bearing MDA-MB-231 xenograft by CTZ, KCZ and ICZ through the accumulation of LC 3B in tumor xenograft. In conclusion, imidazole antifungal drugs induced apoptosis, cell cycle and autophagy by suppressing mTOR through the regulation of PI3K/Akt pathway in human breast cancer cells, suggesting azole antifungal drugs could be a new potential therapeutics for breast cancer.
more목차
I.INTRODUCTION 1
II.METHOD 4
A. Cell lines and Material 4
B. Immunoblot analysis 4
C. Cell cycle assay 5
D. Intracellular ATP assay 5
E. Detection and Quantification of Acidic Vesicular 6
F. Apoptosis assay 6
G. Immunofluorescence 6
H. In vivo Tumor Xenograft Study 7
I. Tissue fractionation 7
J. Statistical analysis 7
III. RESULTS 9
A. Azole antifungal drugs induced apoptosis in human breast cancer cell line 9
B. Imidazole of azole drugs induced G0/G1 arrest in human breast cancer cell line 11
C. Azole antifungal drugs induced autophagy in human breast cancer cell line 14
D. PI3K-Akt inhibition is an important mechanism in causing Azole-induced autophagy in human breast cancer cell lines 18
E. LKB1- AMPK pathway is causing Azole-induced autophagy in human breast cancer cell lines 20
F. Azole antifungal drugs inhibited tumor growth though induced autophagy in a xenograft model 22
IV. DISCUSSION 24
V. CONCLUSION 27
REFERENCES 29
KOREAN ABSTRACT 33
