The Characteristics of Senescent Tumor Cells in Colorectal Cancer
- 주제(키워드) Cellular Senescence , Senescent Tumor Cells , Cancer Evolution , Spatial Evolution
- 주제(DDC) 570
- 발행기관 아주대학교 일반대학원
- 지도교수 Tae Jun Park
- 발행년도 2024
- 학위수여년월 2024. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000033285
- 본문언어 한국어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
목차
Ⅰ. INTRODUCTION 1
1. Cellular Senescence and Cancer 1
2. Detection of Senescent Tumor Cells in Vitro and in Vivo 2
3. Cancer Promoting Features of Senescent Cells in cancer tissues 3
4. Cellular Senescence and Spatial Evolution of Cancer 5
Ⅱ. MATERIALS & METHODS 7
1. IHC analysis of CRC tissues 7
2. IHC Imaging Analysis 8
3. Visium Spatial Transcriptomics Analysis 8
4. Single-cell RNA-sequencing (scRNA-seq) 9
5. Microdissection-assisted RNA-sequencing (mdaRNA-seq) 10
6. Cell Culture and Induction of Cellular Senescence in Vitro 10
7. SA-β-Gal Staining 11
8. SW620 Bulk RNA-sequencing 11
9. Immunoblotting 12
10. Transfection of Vectors and siRNAs 12
11. TCGA Data Set Analysis 13
12. Real-time Polymerase Chain Reaction (RT-PCR) 13
13. Cell Invasion Assay 14
14. Methylation-specific PCR (MSP) 14
15. Estimation of ROS Levels in vivo 14
16. Lentivirus preparation 15
17. Statistical analysis 15
Ⅲ. RESULT 16
Part I: Finding Novel Markers for Senescent Tumor Cells 16
1. Detecting Senescent Tumor Cells using Conventional Senescence Markers 16
2. scRNA-seq of CRC Dataset GSE166555 19
3. scRNA-seq Analysis of CDKN2A-positive Cells Using GSEA 21
4. In vivo validation of p15INK4B as a marker for senescent tumor cells in CRC 23
5. In vitro validation of p15INK4B as a marker for senescent tumor cells in CRC 25
6. Cytokeratin 7 as a marker for senescent tumor cells in CRC 28
Part II: Cancer-promoting features of senescent tumor cells 30
1. Location of senescent tumor cells is in the CRC tissue 30
2. Spatial transcriptomics analysis of CRC tissues using FFPE samples 39
3. Senescent Tumor Cells at the Invasive Front Exhibit Distinctive Phenotype 42
4. Senescent Cells Evolve from Non-senescent Tumor Cells 52
5. Senescence-related Epigenetic Reprogramming in Cancer Evolution 58
6. Senescent Tumor Cells Express Unique Genes to Invade 71
7. LAMC2-EGFR-MMP7 Signaling Axis in Type II Senescent Tumor Cells 81
8. Type II Senescent Tumor Cells are Related to Lymph Node Metastasis 90
Ⅳ. DISCUSSION 98
1. Detection of Senescent Tumor Cells and Limitations 98
2. Cellular Senescence and Cancer Evolution 101
Ⅴ. REFERENCES 109
