Apoptosis Effect of Pancreatic Cancer Cells by Combining Eugenol and TRAIL
Eugenol 과 TRAIL 의 병용처리에 의한 췌장암 세포의 사멸 효과
- 주제(키워드) 도움말 TNF-related apoptosis-inducing ligand , Eugenol , death receptor 5 , endoplasmic reticulum stress , ROS , siRNA
- 발행기관 강릉원주대학교 일반대학원
- 지도교수 도움말 이대희
- 발행년도 2024
- 학위수여년월 2024.02
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 웰니스바이오산업학과
- 세부분야 해당없음
- 실제URI http://www.dcollection.net/handler/kangnung/000000011739
- UCI I804:42001-000000011739
- 본문언어 영어
초록/요약 도움말
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is not toxic to normal cells and is preferentially cytotoxic to cancer cells. However, upon continuous treatment with TRAIL, various carcinomas were resistant. Therefore, there is a need to develop sensitizers that can increase TRAIL susceptibility. Eugenol, the main component of clove oil extracted from clove flower buds, is known to have anti-cancer effects in various studies. In this study, we investigated whether the combination of Eugenol and TRAIL can induce apoptosis in pancreatic cancer cells. When pancreatic cancer cells were treated in combination with Eugenol and TRAIL, it was confirmed that apoptosis was induced compared to when treated alone. Eugenol induced the death receptor DR5 in cancer cells, inhibited DR4 in normal cells, and increased the anti-apoptotic protein FLIP and the tumor suppressor protein p53. It also induced the production of ROS and increased the levels of ER stress-related proteins, including phosphorylated PERK and CHOP. Moreover, ER stress-induced CHOP significantly enhanced DR5 expression, and CHOP knockdown reduced the combined effect of Eugenol and TRAIL. This study demonstrated that Eugenol induces ER stress by overexpressing ROS and enhances TRAIL-induced apoptosis by upregulating DR5 through the PERK-CHOP pathway. In addition, it was demonstrated that it enhanced ER stress by inducing p53, a tumor suppressor protein, and downregulated the expression of the anti-apoptotic protein FLIP. Therefore, we propose that Eugenol is a novel sensitizer that increases sensitivity to TRAIL.
more목차 도움말
Ⅰ. Introduction.........................................................................................................10
Ⅱ. Materials and Methods ...................................................................................13
2.1. Cell culture .............................................................................................13
2.2. Reagent and Antibodies ......................................................................13
2.3. WST-1 assay.............................................................................................14
2.4. Crystal violet assay..................................................................................14
2.5. Colony formation assay...........................................................................14
2.6. Wound healing assay...............................................................................15
2.7. Flow Cytometry Analysis........................................................................15
2.8. Western Blotting......................................................................................15
2.9. Caspase 3/7 assay.....................................................................................16
2.10. ROS measurement.................................................................................16
2.11. Transfection (siRNA).............................................................................17
2.12. Statistical analysis..................................................................................17
Ⅲ. Results .............................................................................................................18
3.1. Effects of TRAIL and Eugenol on human pancreatic cancer cell lines
..........................................................................................................................18
3.2. Eugenol at subtoxic (less toxic) concentrations enhances TRAIL
induced Cell Death in pancreatic cancer cells..............................................22
3.3. Combination of Eugenol and TRAIL induces apoptosis in pancreatic
cancer cells......................................................................................................28
3.4. Eugenol increases TRAIL sensitivity by upregulating DR5................32
3.5. Eugenol induces ROS and activates the ER stress pathway................36
3.6. Eugenol -induced enhanced apoptosis by DR5 and CHOP
upregulation is ROS-dependent....................................................................40
3.7. Effects of Eugenol on the levels of anti-apoptotic proteins, and DRs
of two human pancreatic cancer cell lines....................................................45
Ⅳ. Conclusion .......................................................................................................49
Ⅴ. References .........................................................................................................52
