인터루킨-6 를 표적으로 하여 위암에서 기질유발 항암제 저항성을 극복하기 위한 연구
Targeting interleukin-6 as a strategy to overcome stroma-induced resistance to chemotherapy in gastric carcinomas
- 주제(키워드) 위암 , 암미세환경 , 인터루킨-6 , 암 관련 섬유모세포 , 항암제 저항성 , 토실리주맙
- 발행기관 아주대학교
- 지도교수 허훈
- 발행년도 2018
- 학위수여년월 2018. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000027241
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
In solid tumors such as gastric cancer (GC), the stroma plays a crucial role in decreasing responsiveness to chemotherapy. However, specific targets have not yet been identified to inhibit the interaction between stroma and cancer cells to inhibit stroma-induced chemotherapeutic resistance. The present study aimed to determine whether cancer-associated fibroblasts (CAFs), a major component of tumor stroma, confer chemotherapeutic resistance in GC and what drives this interaction resulting in chemotherapeutic resistance in in vitro and in vivo experimental models. Transcriptome analysis and validation revealed that interleukin-6 (IL-6) is a CAF-specific secretory protein that activates GC cells via paracrine signaling. Furthermore, the IL-6/Jak1/STAT3 signal transduction pathway in CAFs conferred stroma-induced resistance to chemotherapy in GC cells. When monoclonal anti-IL-6R antibody (tocilizumab) was added to chemotherapeutic agents, CAF-induced inhibition of apoptosis was notably abrogated in GC cells. Furthermore, tocilizumab had the same synergistic effects during chemotherapy on GC xenograft tumors in animal models. Clinical data revealed that IL-6 was prominently expressed in the stromal portion in GC tissues; consequently, upregulation of stroma-related genes, including IL-6, in GC tissues was correlated with poor responsiveness to chemotherapy. Our data provide plausible evidence for crosstalk between GC cells and CAFs, wherein IL-6 plays a key role in initiating chemotherapeutic resistance. The present findings suggest the use of IL-6 inhibitor as therapeutic agent to enhance responsiveness to chemotherapy in GCs.
more목차
I. INTRODUCTION 1
II. MATERIALS AND METHODS 3
1. Cell lines and cell culture 3
2. Isolation and culturing of fibroblasts 3
3. Co-culture with CAFs or NAFs 3
4. Western blot 4
5. Transcriptome analysis and quantitative real-time PCR (qRT-PCR) 5
6. Reverse transcriptase polymerase chain reaction (RT-PCR) 6
7. Enzyme-linked immunosorbent assay 7
8. Cell viability assay 7
9. Immunohistochemical staining 8
10. Generation of short hairpin (sh) RNA for IL-6 (shIL-6) 8
11. Animal model study 9
12. TCGA data 10
13. Gene expression analysis in pretreated biopsied tissues 10
14. Statistical analysis 11
III. RESULTS 12
1. Fibroblasts reduce responsiveness to 5-FU in GC cells 12
2. CAF-secreted IL-6 activated the JAK1/STAT3 signal transduction pathway in
GC cell lines 20
3. Inhibition of IL-6/Jak1/STAT3 axis suppresses the drug resistance in GC cell
lines 33
4. Tocilizumab reverses the effect of CAF-induced chemotherapeutic in the
xenograft mouse model for GCs 41
5. Stroma-related genes including IL-6 in biopsied tissues reduce
responsiveness to chemotherapy in GCs 47
IV. DISCUSSION 56
V. REFERENCES 63
국문요약 69
