인간 일차 간세포에서 FoxM1 발현을 통한 AKT 활성으로 B-RafV600E에 의해 유도된 노화 극복 연구
Abrogation of B-RafV600E induced senescence by AKT activation via FoxM1 expression in primary human hepatocytes
- 주제(키워드) Hepatocellular carcinoma; HCC; Hepatocarcinogenesis; Primary human hepatocytes; B-RafV600E; PTEN; FoxM1
- 발행기관 아주대학교
- 지도교수 박태준
- 발행년도 2017
- 학위수여년월 2017. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000024361
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world. Hepatocarcinogenesis is a stepwise process and multiple genes alteration are involved such as activation of oncogenes, inactivation of tumor suppressor genes, activation of growth factor and their cognitive receptors, reactivation of developmental pathways and activation of cell cycle regulator. In general, MAPK and the PIK3CA signalling pathways have a key role in cell proliferation and survival and their oncogenic activation deeply contributes to pathogenesis of different solid tumors. B-Raf mutation has been reported in 61% of melanoma, 53% of papillary thyroid cancer and 11.5% of colorectal cancer patients. In case of hepatoma, 23% of B-RafV600E mutation has been reported. Furthermore, PI3K pathway activation was found about 40% of HCC cases. Therefore, in this study, we evaluated the role of B-RafV600E and PI3K pathway signaling in hepatocarcinogenesis. Overexpression of B-RafV600E in primary human hepatocytes caused cell growth arrest and increased senescence-associated beta-galactosidase (SA-β-gal) staining. Furthermore, we found that shPTEN induced cellular senescence. We further investigated that shPTEN and B-RafV600E co-infection effect in hepatocarcinogenesis and found that overcome of senescence was observed. At the mechanism level, B-RafV600E infection decreased FoxM1 expression. However, co-infection increased FoxM1 expression. Furthermore, B-RafV600E and FoxM1 co-infected hepatocytes also showed overcome of B-RafV600E induced senescence.
more목차
I. INTRODUCTION ················································································· 1
II. MATERIALS AND METHODS ······························································ 4
1. Cell line and Culture conditions ····························································· 4
2. Plasmids and Lentivirus Preparation ························································ 4
3. RNA interference ·············································································· 4
4. Immunoblotting ················································································ 5
5. Quantitative Real-time RT–PCR ···························································· 5
6. Senescence-Associated β-Galactosidase staining ········································· 6
7. Soft agar colony forming assay ······························································ 6
8. Statistical Analysis ············································································· 6
III. RESULTS························································································· 7
1. B-RafV600E induced senescence in primary human hepatocytes. ····················· 7
2. Downregulation of PTEN in primary human hepatocytes induced senescence. ···· 14
3. Senescence induced by B-RafV600E can be overcome with shPTEN co- infection. ······························································································ 18
4. B-RafV600E and FoxM1 co-infected hepatocytes overcome the B-RafV600E induced senescence. ················································································· 24
IV. DISCUSSION ··················································································· 32
V. REFERENCES ·················································································· 34
국문요약 ···························································································· 37
