상피-중간엽 이행 유도에 의한 유방암세포의 지질대사 조절인자 분석
Analysis of lipid metabolism regulators in breast cancer cells after induction of the Epithelial to Mesenchymal Transition
- 주제(키워드) Metastasis , EMT , Lipid metabolism , Fatty acid , PPAR-γ , Breast Cancer
- 발행기관 아주대학교
- 지도교수 김유선
- 발행년도 2015
- 학위수여년월 2015. 8
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000020516
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The process of tumor metastasis is often enabled by the epithelial-mesenchymal transition (EMT), a key developmental program that is often activated during cancer invasion and metastasis. During the EMT process, cells undergo phenotypic changes, lose of E-cadherin expression and acquire invasive properties that allow them to migrate through the extracellular matrix. EMT inducers or regulators have been shown to induce cancer cells to form populations with cancer stem cell-like characteristics, providing them with therapeutic resistance and conferred tumor recurrence. Although metabolism plays a fundamental role in essentially every function of a cell, little is known about how the EMT programme involves in regulating cell’s metabolism contributes to the morphological and molecular changes. An early and universal feature of tumors is the activation of lipid metabolism, and it is a central hallmark of many cancers including prostate and breast cancer. To understand lipid metabolism in cancer, we used LC-MS to identify lipid construct. Interestingly, after induction of EMT phenotype, cells show global fatty acid metabolism changes. We found that peroxisome proliferator activated receptor-gamma (PPAR-γ), a master transcriptional regulator of adipogenesis, was downregulated during EMT process. Downregulation of PPAR-γ in breast cancer cells enhances cell migration, colony formation, and chemotherapy resistance indicating PPAR-γ has critical role in breast cancer metastasis.
more목차
Ⅰ. INTRODUCTION 1
Ⅱ. MATERIALS AND METHODS 3
A. Reagents 3
B. Cell cuture 3
C. Mammosphere culture 3
D. LC-MS based cellular lipid profiling 4
E. GC-MS based cellular esterified fatty acid profiling 5
F. Westernblot analysis 6
G. Lentiviral shRNA experiments 6 H. Cytotoxicity assay 7
I. Reverse Transcription-PCR 7
J. Colony formation assay 7
K. Wound Healing assay 9
L. Total RNA extraction and gene expression profiling 9 M. Public microarray data analysis 10
N. Gene set enrichment analysis 10
O. Kaplan-Meier analysis 10
Ⅲ. RESULTS 11
A. Exchange of lipid metabolism-related genes after EMT induction in MCF-7 cells 11
B. Differentially regulated lipids between adherent and sphere form of MCF-7 cells 17
C. Precise alteration of fatty acid metabolism during EMT induction in cancer cells 21
D. Various EMT induction conditions induce alteration of lipid metabolism-related genes 25
E. The decrease of PUFA biosynthesis mediates malignant phenotype of cancer cells via
inhibition of PPAR-γ expression 27
Ⅳ. DISCUSSION 35
Ⅴ. CONCLUSION 37
Ⅵ. REFERENCES 38
국문요약 44
