기질 세포에 의해 유도되는 위암 복막 전이에서 discoidin domain receptor 1의 역할에 대한 연구
Inhibition of discoidin domain receptor 1 prevents stroma-induced peritoneal metastasis in gastric carcinomas
- 주제(키워드) Gastric carcinomas , Peritoneal metastasis , Discoidin domain receptor , Cancer-associated fibroblasts
- 발행기관 아주대학교
- 지도교수 허훈
- 발행년도 2017
- 학위수여년월 2017. 8
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000025572
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Although the interactions between cancer cells and the tumor stroma are known to play a role in the peritoneal metastasis of gastric carcinomas (GCs), effective targeting agents that block these interactions have never been reported. Discoidin domain receptor 1 (DDR1) is activated by triple-helix collagens, which are major components of tumor stroma; thus, DDR1 might have a key role in the communication between cancer cells and stroma. The aim of this study was to investigate the effect of DDR1 inhibition on stroma-induced peritoneal metastasis in GCs. Methods: We analyzed by immunohistochemistry the correlation between DDR1 expression and the pattern of recurrence in GC tissues from a cohort established in a previous study. We also co-cultured human GC cell lines with gastric cancer-associated fibroblasts (CAFs), and investigated DDR1 expression and activation by western blotting. We evaluated the CAF-induced tumorigenic ability of GC cell lines and the effects of a DDR1-specific inhibitor in organotypic cultures and a peritoneal seeding xenograft animal model. Results: The expression of DDR1 in gastric cancer tissues was significantly positively associated with early recurrence (p = 0.043) and a high incidence of peritoneal recurrence (p = 0.036). We confirmed that co-culture with CAFs elevated DDR1 protein expression in GC cell lines. CAFs also enhanced GC cell line spheroid formation in organotypic cultures in a tumor cell DDR1-dependent manner. Co-implantation of CAFs with GC cells enhanced peritoneal tumor formation in vivo, an effect that was sensitive to pharmacologic inhibition of DDR1. Conclusion: Our findings suggest that the CAF-induced elevation of DDR1 in GC cells enhances their peritoneal tumorigenesis, and that the inhibition of DDR1 signaling is an attractive strategy for the treatment of GC peritoneal metastasis.
more목차
I. INTRODUCTION 1
II. MATERIALS AND METHODS 3
1. Human samples and immunohistochemical staining 3
2. Cell lines and chemicals 3
3. CRISPR/Cas9-mediated knockout of DDR1 4
4. Western blot 5
5. Cell viability test 5
6. RT-PCR 6
7. Organotypic culture and immunocytochemical staining 6
8. Peritoneal xenograft mouse models 6
9. Statistical analysis 7
III. RESULTS 8
1. DDR1 expression correlates with peritoneal recurrence in primary GC tissues 8
2. CAFs enhance the peritoneal tumorigenic potential of GCs 11
3. CAFs-induced upregulation of DDR1 in GC cell lines 14
4. DDR1 inhibition using the CRISPR/Cas9 system suppresses the effects of CAFs 20
5. Blocking DDR1 signaling through 7rh represses the effects of CAFs on GCs 22
IV. DISCUSSION 30
V. REFERENCES 35
국문요약 42
