Title Page
Abstract
Contents
List of abbreviation 19
Background 22
1. Biomarkers in cancer research 22
2. Nonsteroidal anti-inflammatory drug (NSAID)-activated gene-1 (NAG-1, GDF15) 25
3. Thrombospondin-1 (TSP-1) 33
4. Alternative splicing in cancer 39
5. Thyroid cancer 42
6. Canine mammary tumor and canine osteosarcoma 48
CHAPTER Ⅰ. Quercetin induces anticancer activity by upregulating pro-NAG-1/GDF15 in differentiated thyroid cancer cells 49
Abstract 50
1.1. Introduction 51
1.2. Materials and Methods 54
1.3. Results 62
1.4. Discussion 82
CHAPTER Ⅱ. Novel thrombospondin-1 transcript exhibits distinctive expression and activity in thyroid tumorigenesis 87
Abstract 88
2.1. Introduction 89
2.2. Materials and Methods 92
2.3. Results 107
2.4. Discussion 131
CHAPTER Ⅲ. Novel TSP-1 variants as diagnostic biomarkers and therapeutic targets in canine mammary tumors 137
Abstract 138
3.1. Introduction 139
3.2. Materials and Methods 142
3.3. Results 148
3.4. Discussion 169
General conclusion 173
Bibliography 177
국문초록 195
Background 18
Table B-1. Ten leading cancer types for the new cancer cases and percentage by sex, Korea, 2020 46
Chapter Ⅰ 18
Table 1-1. Demographics for patients with thyroid cancer 68
Chapter Ⅱ 18
Table 2-1. Demographics for patients with thyroid cancer 93
Table 2-2. Demographics for patients with thyroid cancer (RT-qPCR) 94
Table 2-3. Primers used in chapter II. 99
Chapter Ⅲ 18
Table 3-1. Primers used in chapter III 147
Table 3-2. Demographics for patients with canine mammary tumor 154
Background 14
Figure B-1. Classification of biomarkers based on its main clinical application 24
Figure B-2. The generation process and structure of pro- and mature NAG-1 30
Figure B-3. Pleiotropic effects of GDF15 in malignancies: GDF15 regulates tumor metastasis, cachexia, and immune infiltration 31
Figure B-4. Boxplot for NAG-1 expression profile across cancer experiments (GPL570 platform) 32
Figure B-5. Structure and biological functions of TSP-1 37
Figure B-6. Boxplot for TSP-1 expression profile across cancer experiments (GPL570 platform) 38
Figure B-7. Illustration of the seven types of alternative splicing 41
Figure B-8. Ten leading cancer types for the estimated new cancer cases and deaths by sex, United States, 2023 47
Chapter Ⅰ 15
Figure 1-1. Antibody array using human thyroid tissues 64
Figure 1-2. NAG-1/GDF-15 concentration was increased by age 66
Figure 1-3. NAG-1 expression in various types of cancer cell lines 70
Figure 1-4. Pro-NAG-1/GDF15 expression in the presence of several anticancer compounds 72
Figure 1-5. Quercetin induced apoptosis and cell cycle arrest in BCPAP cells 75
Figure 1-6. Quercetin increased NAG-1 promoter activity 79
Figure 1-7. Proposed mechanism by which pro-NAG-1 plays an anti- tumorigenic role in thyroid cancer 81
Chapter Ⅱ 15
Figure 2-1. Thrombospondin 1 (TSP-1) was selected as a strong candidate diagnostic biomarker for thyroid cancer 109
Figure 2-2. TSP1V expression in conditioned medium (CM) and cell lysates (CL) depends on serum levels 113
Figure 2-3. Overexpressing TSP1V functionally affects thyroid cancer progression 117
Figure 2-4. Intron-retained TSP1V expression is affected by compound treatment 122
Figure 2-5. Mechanisms of IR induced by compounds (T-C and SS) 126
Figure 2-6. Differential expression of TSP1W and TSP1V in follicular thyroid cancer 130
Chapter Ⅲ 16
Figure 3-1. Thrombospondin 1 (TSP-1) expression in breast cancer and osteosarcoma patients 150
Figure 3-2. Identification of transcriptional variants of dTSP-1 in canine cell lines 155
Figure 3-3. Protein similarity of TSP-1 among species 156
Figure 3-4. Anti-cancer compound affects cell proliferation in CF41.Mg and D17 cell lines (Complete medium) 158
Figure 3-5. Anti-cancer compound affects cell proliferation in CF41.Mg and D17 cell lines (Serum free medium) 159
Figure 3-6. Intron-retained dTSP1V expression is affected by compound treatment in CMT and COS 161
Figure 3-7. Intron-retained TSP1V expression is affected by compound treatment in HBC and HOS 164
Figure 3-8. Intron-retained TSP1V expression induced by compound treatment in MDA-MB-231 166
Figure 3-9. dTSP-1 expression in normal and tumor tissues from CMT patients 168