Isoalantolactone Suppresses Tumor Progression by regulating Hippo-YAP Signaling in Cholangiocarcinoma cell
- 주제(키워드) Cancer
- 주제(DDC) 570
- 발행기관 아주대학교 일반대학원
- 지도교수 Jung-Soon Mo
- 발행년도 2024
- 학위수여년월 2024. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000033733
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Cholangiocarcinoma (CCA), which originates from bile duct epithelial cells, is a malignancy with poor prognosis owing to limited treatment. Here, we report the anti- cancer effect and mechanism of isoalanthoactone (IALT) through the Hippo pathway in CCA cells. In our study, we initially identified the cytotoxic effect of IALT, which inhibits cell proliferation and induces apoptosis in SNU478 cells, a CCA cell line. This effect stems from the regulation of MST1/2 and LATS1/2 by IALT, inducing phosphorylation of YAP, an oncogene of CCA, and inhibiting its activity. Therefore, the translocation to the nucleus of the YAP was inhibited, disrupting interaction with the TEAD transcription factor, thereby reducing the transcription of the YAP-target genes, CYR61 and CTGF. Furthermore, we found that IALT inhibits cell growth, cell migration, and CCA tumor formation in xenograft mouse model. Through LATS1/2-deficient cells, we confirmed the importance of LATS1/2 in the mechanism of IALT by showing a reduction in the effectiveness of IALT. Our data demonstrated that IALT functions as an anti-cancer drug by inhibiting YAP activity in CCA tumor progression. These findings propose a potential therapeutic agent for the treatment of CCA and highlight the importance of YAP-target therapies. Keywords: cholangiocarcinoma, isoalantolactone, Hippo pathway, YAP, LATS1/2, MST1/2, TEAD
more목차
I. Introduction 1
II. Material and Methods 5
1. Antibodies 5
2. Chemicals 5
3. Cell culture and transfection 5
4. CRISPR/Cas9 system 6
5. Retroviral infection and generation of stable cell lines 6
6. Cell lysis and Western blotting 7
7. MTT assay 7
8. Cell apoptosis assay 8
9. Immunoprecipitation (IP) 8
10. Immunocytochemistry 9
11. RNA isolation and qRT-PCR (quantitative real-time polymerase chain reaction) 9
12. Wound healing assay 10
13. Clonal growth assay 11
14. TA cloning 11
15. Tumor isolation from mouse 12
III. Results 13
1. Cytotoxic effect of IALT induces apoptosis in SNU478 cells 13
2. IALT regulates YAP activity via Hippo pathway 16
3. Absence of LATS1/2 counteracts the effect of IALT in regulating YAP 21
4. IALT inhibits Cell growth, Cell Migration and Tumorigenesis by controlling YAP activity 30
IV. Discussion 37
V. References 40
국문 초록 43
