Ivermectin kills cancer cells via catastrophic changes in the endoplasmic reticulum structure
Ivermectin kills cancer cells via catastrophic changes in the endoplasmic reticulum structure
- 주제(키워드) Ivermectin , ER , Calcium , cancer , Chloride , Paraptosis
- 발행기관 아주대학교
- 지도교수 최경숙
- 발행년도 2019
- 학위수여년월 2019. 8
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000029372
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Ivermectin (IVM), an antiparasitic drug, is now considered a strong candidate for repurposing as an anticancer drug. IVM has been known to induce apoptosis or autophagy in several cancer cells, but the underlying mechanisms to explain its anticancer effects still remain unclear. In this study, we found that induces dramatic alterations of the endoplasmic reticulum (ER) structure, including reorganization, vacuolation, and subsequent permeabilization of the ER, prior to cancer cell death. In addition, IVM increases ER stress, and pretreatment with cycloheximide, a protein synthesis inhibitor, markedly blocked IVM-induced ER stress, vacuolation and permeabilization of the ER, and cell death, but not ER reorganization. Furthermore, IVM induces the activation of ERK and p38, imbalance of ions, including Ca2+ and Cl-, and inhibition of these signals effectively attenuated IVM-induced vacuolation and permeabilization of the ER and subsequent cell death. Taken together, these results suggest that IVM-induced catastrophic changes in the ER, including the vacuolation and permeabilization of the ER, critically contribute to the anti-cancer effect of IVM. ER stress, imbalance of Ca2+ and Cl-, and activation of MAPK play crucial roles in this process.
more목차
I. INTRODUCTION 1
II. MATERIALS AND METHODS 11
A. Chemicals and antibodies 11
B. Cell culture 12
C. Measurement of cellular viability 12
D. Immunoblot analyses 13
E. Morphological examination of the ER and mitochondria 13
F. Immunofluorescence microscopy 14
G. Transmission electron microscopy 14
H. Measurement of intracellular Ca2+ levels 14
I. Measurement of intracellular Cl- levels 15
J. Measurement of intracellular reactive oxygen species levels 15
K. Measurement of cellular oxygen consumption Rate 15
L. ATP measurement 16
M. Statistical analysis 16
III. RESULTS 17
1. Ivermectin (IVM) effectively induces cell death in various cancer cells 17
2. IVM induces catastrophic changes in the endoplasmic reticulum structure 23
3. ER reorganization is associated with lipid 30
4. IVM induces paraptosis in MDA-MB 435S 34
5. Increase in intracellular Ca2+ critically contributes to IVM-induced paraptosis 41
6. Imbalance in intracellular chloride may be involved in IVM-induced paraptosis 47
IV. DISCUSSION 53
V. REFERENCES 67
