New target for Cetuximab Resistance For Colorectal Cancer
대장암 항암제 세툭시맙의 새로운 표적
- 주제(키워드) 도움말 Colorectal Cancer , Cetuximab Resistance , S100A4 Overexpression , Neferine , Drug Resistance Mechanisms , SW48 cells , Targeted Therapy , Cancer Pharmacology.
- 발행기관 강릉원주대학교 일반대학원
- 지도교수 도움말 이대희
- 발행년도 2024
- 학위수여년월 2024. 2
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 웰니스바이오산업학과
- 세부분야 해당없음
- 실제URI http://www.dcollection.net/handler/kangnung/000000011713
- UCI I804:42001-000000011713
- 본문언어 영어
초록/요약 도움말
Colorectal cancer is still a major cause of death globally. Although targeted therapies, such as cetuximab, are changing the treatment of advanced or metastatic colorectal cancer, drug resistance remains a significant challenge. This study investigates the correlation between cetuximab resistance and S100A4 expression in patients with RAS-wild type metastatic colorectal cancer and exploring the potential to overcome this resistance through the combined treatment of Neferine and Cetuximab. The study aimed to induce cetuximab-resistant SW48R cell lines from RAS-wild type SW48 colorectal cancer cells through repeated cetuximab treatments. Microarray analysis identified a relationship between these resistant cells and the S100A4 gene, which is connected to the Hypoxia pathway. Protein expression analysis across various colorectal cancer cells revealed an increase in S100A4 expression alongside cetuximab resistance, particularly in SW48R cells, indicating increased S100A4 expression through the PI3K/AKT/GSK-3β pathway. Further validation came from Immunohistochemistry (IHC) of colorectal cancer tissues collected from patients at Guro Hospital and subsequent protein expression experiments, both confirming increased expression of S100A4. Treatment with Recombinant S100A4 protein (rS100A4) activated the PI3K/AKT/GSK-3β pathway, elevating S100A4 levels, and resulted in reduced cetuximab sensitivity in SW48R cells, as evident in cell viability and apoptosis assays. Drug screening efforts to overcome cetuximab resistance identified Neferine as an effective compound, demonstrating cell death in cetuximab-resistant cells and decreasing PI3K/AKT/GSK-3β pathway activity and S100A4 protein expression. The combined treatment of Cetuximab and Neferine proved more effective than Cetuximab alone. This research underscores the correlation between overexpression of S100A4 and cetuximab resistance in colorectal cancer, proposing strategies targeting S100A4 to overcome resistance. This approach highlights the potential for improved treatment outcomes in colorectal cancer patients and underscores the importance of strategically targeting S100A4 in drug-resistant colorectal cancer.
more목차 도움말
ABSTRACT 08
Ⅰ. Introduction 12
Ⅱ. Materials and Methods 15
2.1. Antibodies and reagents 15
2.2. Cell culture and Drug treatments 15
2.3. Microarray analysis for cetuximab resistance pathways 16
2.4. Patient data collection and ethical approval for colorectal cancer study at Gro hospital 16
2.5. Cell proliferation assay 17 2.6. Annexin V-FITC/PI staining assay 17
2.7. Colony-forming assay 18
2.8. Western blot 18
2.9. Statistical analysis 19
Ⅲ. Experimental Results
3.1. Comprehensive Gene Expression Analysis in SW48 and Cetuximab-Resistant SW48R Colorectal Cancer Cells 20
3.2. Analysis of S100A4 Expression and PI3K/AKT/GSK-3β Pathway Activation in Various Colorectal Cancer Cells and Their Cetuximab-Resistant Counterparts 25
3.3. Effects of recombinant S100A4 protein on SW48 cell viability and pathway activation 30
3.4. Evaluation of cetuximab resistance in SW48R and rS100A4 treated SW48 cells 34
3.5. Evaluation of Cetuximab Resistance in SW48R and rS100A4-Treated SW48 Cells Immunohistochemistry and Western Blot Analysis of S100A4 in Human Colorectal Cancer Tissues 40
3.6. Screening of Nelumbo nucifera components for S100A4 inhibition in SW48R cells 45
3.7. Overcoming cetuximab resistance in SW48 and SW48R cells by combined treatment with Neferine 51
Ⅳ. Conclusion 58
Ⅴ. References 60
