커큐민의 JAK/STAT3 신호 전달 경로의 억제를 통한 위암의 암 관련 섬유 아세포 유래 항암제 저항성 억제에 대한 연구
Curcumin inhibits cancer-associated fibroblast-derived chemoresistance of gastric cancer through suppression of the JAK/STAT3 signaling pathway
- 주제(키워드) Gastric cancer , Cancer-associated fibroblasts , Chemoresistance , JAK/STAT3 signal transduction pathway , Curcumin
- 발행기관 아주대학교
- 지도교수 허훈
- 발행년도 2020
- 학위수여년월 2020. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000029609
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Introduction: Gastric cancer (GC) is one of the most common malignant tumors worldwide. However, very few studies have been successful in the discovery and clinical application of new anticancer drugs to increase the survival rate of gastric cancer patients. Cancer-associated fibroblasts (CAFs) are a major component of tumor stroma and their effect on the progression of GC has recently been demonstrated. Cytokines or growth factors, which are ligands that activate the JAK/STAT3 signaling pathway, are secreted from stromal cells such as fibroblasts in the tumor microenvironment, and gastric cancer, which has a large amount of fibroblasts in gastric cancer, exhibits a poor clinical prognosis. In the previous study, we demonstrated that, CAFs-derived chemoresistance of gastric cancer through the IL-6/JAK/STAT3 signaling pathway. The present study aims to investigate the JAK/STAT3 signaling pathway control mechanism of the gastric cancer cell by CAFs, and suggests curcumin, an inhibitor for STAT3, as a potential candidate for GC treatment. Methods: The cell viability assay and flow cytometry were utilized for analyzing GC cell growth and apoptosis in CAFs-conditioned media (CM) when treated with 5-fluorouracil (5-FU). Secretome analysis was performed to evaluate levels of several secreted proteins from CAFs co-cultured with GC cells. Immunoblotting was performed to examine whether curcumin suppressed JAK2/STAT3 signaling pathway. We also co-cultured human GC cells with CAFs and investigated phosphorylation of JAK2 and STAT3 by western blotting. The effect of curcumin, inhibitor of STAT3, on the CAFs-induced chemoresistance of GC cells was also investigated. Results: The cell viability assay revealed that the CM collected from CAFs further weakened the response to chemotherapeutic agents like 5-FU and cisplatin in GC cell lines. Secretome analysis showed that levels of several secreted proteins, e.g., IL-6, IL-8, and CCL2, increased in CM from CAFs co-cultured with cancer cells (CAFs-CM), when compared with those in CM from cancer cells only. Western blot analysis revealed that CAFs could induce the activation of the JAK2/STAT3 signaling pathway in cancer cells. Cell viability assays and flow cytometry revealed that curcumin significantly decreased the chemoprotective capacity of CAFs-CM. Conclusion: These data strongly suggest that suppression of the JAK/STAT3 signal transduction pathway counteracts the effect of CAFs-induced chemotherapeutic resistance in GC cell lines. We propose that curcumin, inhibitor of STAT3, may be a suitable agent to overcome chemotherapeutic resistance through the inhibition of CAFs-induced activation of the JAK/STAT3 signaling pathway.
more목차
I. INTRODUCTION 1
II. MATERIALS AND METHODS 3
1. Cell culture 3
2. Western blotting 3
3. Cell viability assay 4
4. Secretome and transcriptome analysis 4
5. Flow cytometry analysis 5
6. Xenograft mouse model 5
7. Immunohistochemical staining 6
8. Statistical analysis 7
III. RESULTS 8
1. Cancer-associated fibroblasts reduce chemoresistance to 5-FU in GC cell lines 8
2. Evaluating whether CAFs activate the JAK2/STAT3 signal transduction pathway in GC cell lines 11
3. Determining whether c urcumin inactivated CAFs-induced JAK2/STAT3 signal transduction pathway in GC cell lines 15
4. Curcumin improves the apoptotic effect of chemotherapy on GC cells under co-culture with CAFs 19
5. Combined treatment using curcumin and 5-FU can effectively inhibit tumor growth for CAFs-mixed xenograft tumors 22
IV. DISCUSSION 27
V. REFERENCES 30
국문요약 35
