비소세포폐암에서 microRNA-181a-5p에 의한 당과 지질 대사의 재조정
microRNA 181a-5p Reprogramed Glucose and Lipid Metabolism in Non- Small Cell Lung Cancer
- 주제(키워드) microRNA , Aerobic glycolysis , lipid metabolism , cancer metabolism , Lung cancer
- 발행기관 아주대학교
- 지도교수 임상현
- 발행년도 2020
- 학위수여년월 2020. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의학과
- 실제URI http://www.dcollection.net/handler/ajou/000000029522
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
microRNAs play an important role in cancer metabolisms. miRNA-181a-5p has decreased expression, particularly in non small cell lung cancer tissues and mesenchymal like lung cancer cells. Epithelial-mesenchymal transition mechanisms and cancer metabolism are regulated by the same signaling pathway. Functional analysis was performed to identify the functional role of miRNA-181a-5p in cancer metabolism and to find the target gene. To define the role of miRNA-181a-5p in cancer metabolism, we performed LDH assay, glucose uptake assay, Oil Red O staining, and mitochondrial ATP synthase inhibitor assay. The target gene of miRNA-181a-5p was determined by luciferase reporter assay, qRT-PCR, and western blot analyses. The Functional effect of miRNA-181a-5p on cancer metastasis was measured by migration and invasion assays. Experimental results showed that overexpression of miRNA-181a-5p reduced aerobic glycolysis and adipocytes and decreased cancer cell invasion and migration. Expression of SIRT1 and ACSL4 was reduced by the binding of miRNA-181a-5p to the 3’-UTR. These results suggest that miRNA-181a-5p might participate in the reprogramming of cancer metabolism.
more목차
I. Introduction 1
A. Introduction 1
1. The relation between Glucose, Lipid metabolism and Cancer cell 1
2. The role of miRNA in glucose and lipid metabolism 1
3. The role of miRNA in Cancer Cell 2
4. miRNA-181a-5p and Non Small Cell Lung Cancer 2
5. Hypothesis 3
II. Subjects 4
A. Materials and Methods 4
1. Cell culture and growth conditions 4
2. Glucose-uptake assay 4
3. Lactate dehydrogenase assay 4
4. Mitochondrial ATP synthase Inhibition assay 4
5. Measurement of extracellular acidification rate (ECAR) after treatment of miRNA-181a-5p 5
6. Lipid Staining 5
7. Invasion and migration assays 5
8. TaqMan miRNA expression assay 6
9. miRNA mimic and siRNA transfection 6
10. qRT-PCR 6
11. Western blot 6
12. Approach to search the potential target genes of miRNA-181a-5p 7
13. Luciferase reporter assays 7
14. Statistical analyses 7
B. Results 8
1. Function of miRNA-181a-5p in aerobic glycolysis 8
2. miRNA-181a-5p reduces total lipid in lung cancer cell 9
3. miRNA-181a-5p inhibits lung cancer cell migration and invasion in vitro 9
4. SIRT1 and ACSL4 transcriptions are repressed by binding of miRNA-181a-5p to the 10
5. Quantitative analysis of miRNA-181a-5p, SIRT1, and ACSL 4 expression in lung cancer cell lines 10
6. miRNA-181a-5p regulates SIRT1 and ACSL4 expression at mRNA and protein levels 11
C. Discussion 11
III. Conclusion 14
