Title Page
Contents
ABSTRACT 8
ABBREVIATIONS 14
1. INTRODUCTION 16
1.1. Diabetes 16
1.2. Cancer 17
1.3. Breast cancer 21
1.4. Diabetes and breast cancer risk 24
1.5. IGF1 and insulin 26
1.6. IGF1R and IR 27
1.7. TRIP-Br1 32
1.8. NEDD4-1 36
2. MATERIALS AND METHODS 37
2.1. Cell lines and reagents. 37
2.2. Suppression or overexpression of TRIP-Br1, NEDD4-1, and IR. 37
2.3. Cell viability analysis. 38
2.4. Western blot analysis. 38
2.5. Quantitative real time polymerase chain reaction (qRT-PCR). 38
2.6. Immunoprecipitation (IP). 39
2.7. Immunofluorescence (IF). 40
2.8. Immunohistochemistry (IHC) analysis. 40
2.9. Animal experiments. 41
2.10. Xenograft study. 41
2.11. Single-cell RNA sequencing analysis. 41
2.12. Statistics analysis. 42
3. RESULTS 44
3.1. The correlation between TRIP-Br1, IR and IGF1R in various cells 44
3.2. The positive effect of TRIP-Br1 on IR expression level 47
3.3. The inhibitor role of TRIP-Br1 on the ubiquitination of IR 50
3.4. The effect of TRIP-Br1 on IR/IGF1R ratio 53
3.5. Enhanced IGF1R expression level by IR knockdown 56
3.6. TRIP-Br1 overexpression reduces the expression of IGF1R 58
3.7. Knocking down of TRIP-Br1 enhances expression of IGF1R 60
3.8. Double knocking down of NEDD4-1/TRIP-Br1 up-regulates IGF1R 64
3.9. TRIP-Br1 directly binds with NEDD4-1 and IGF1R 68
3.10. TRIP-Br1/NEDD4-1 mediates IGF1R degradation via a proteasome/ubiquitination 73
3.11. TRIP-Br1 inhibits the ubiquitination of IGF1R 77
3.12. Enhanced tumor formation is associated with a higher IR/IGF1R ratio resulting from TRIP-Br1 expression 80
3.13. TRIP-Br1 induces a higher IR/IGF1R ratio in insulin-deficient mice mimicking diabetes 84
3.14. TRIP-Br1-mediated higher IR/IGF1R ratio enhances survival rate in breast cancer patients 91
3.15. High TRIP-Br1 enhances survival rate in breast cancer from the patient database 95
3.16. TRIP-Br1 enhances survival rate by stabilizing IR under anti-cancer drug 98
3.17. The effect of TRIP-Br1 on IGF1R/IR downstream signaling pathway 101
4. DISCUSSION 104
5. REFERENCE 109
ABSTRACT IN KOREAN 120
Table 1. Variations in expression of the top 10 upregulated genes in type 1 diabetes across different datasets. 34
Table 2. Patient characteristics are based on a single cell from 11 patients. 92
Figure 1. Death rates by cause in those with and without diabetes in 2001 and 2018 19
Figure 2. Diabetes and cancer 20
Figure 3. Breast cancer. Schematic classification of breast cancer subtypes with relative prognosis and therapy. 23
Figure 4. Shared factors in diabetes and breast cancer risk/progression. 25
Figure 5. Components of the insulin/IGF axis. 29
Figure 6. IR and IGF1R structure. 30
Figure 7. The IR and IGF1R signaling pathway 31
Figure 8. Differentially expressed genes shared between T1D and T2D 35
Figure 9. TRIP-Br1 KO-mice genotyping design 43
Figure 10. The expression level of IGF1R, IR, and TRIP-Br1 in breast normal and cancer cells 46
Figure 11. The impact of TRIP-Br1 on IR expression level 49
Figure 12. The effect of TRIP-Br1 on the ubiquitination of IR 52
Figure 13. The association between TRIP-Br1 and IR/IGF1R ratio 55
Figure 14. The impact of IR on IGF1R expression level in MCF7 and MDA-MB-231 57
Figure 15. Effect of TRIP-Br1 overexpression on the IGF1R 59
Figure 16. Effect of TRIP-Br1 knockdown on the IGF1R 63
Figure 17. The effect of TRIP-Br1/NEDD4-1 on IGF1R expression 67
Figure 18. The TRIP-Br1/NEDD4-1-mediated IGF1R degradation 72
Figure 19. TRIP-Br1/NEDD4-1-mediated IGF1R degradation mainly through the proteasome system 76
Figure 20. The impact of TRIP-Br1 on IGF1R ubiquitination 79
Figure 21. A higher IR/IGF1R ratio resulting from TRIP-Br1 expression enhances the survival rate 83
Figure 22. The effect of TRIP-Br1 on IR/IGF1R ratio in diabetes-mimicking mice 90
Figure 23. TRIP-Br1-mediated IR/IGF1R ratio by single cells analysis 94
Figure 24. Bioinformatics analysis of the relationship between TRIP-Br1 expression, IR/IGF1R ratio, and the survival ratio in three different types... 97
Figure 25. The expression level of IGF1R, IR, and TRIP-Br1 in cells 100
Figure 26. The effect of TRIP-Br1 on IGF1R/IR downstream signaling pathway 103
Figure 27. The cellular levels of ROS in MCF7WT-TRIP-Br1 and MCF7KD-TRIP-Br1 cell lines[이미지참조] 106
Figure 28. Grb10 expression level in MCF7 107
Figure 29. Summary model 108