세포유형에 따른 HECAT5와 M3R의 상호작용을 통한 세포증식 조절
DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | 이명애 | - |
dc.contributor.author | Woo changsung | - |
dc.date.accessioned | 2019-10-21T07:28:18Z | - |
dc.date.available | 2019-10-21T07:28:18Z | - |
dc.date.issued | 2016-08 | - |
dc.identifier.other | 22803 | - |
dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/18892 | - |
dc.description | 학위논문(석사)--아주대학교 일반대학원 :의생명과학과,2016. 8 | - |
dc.description.tableofcontents | Ⅰ. INTRODUCTION ···································································································· 1 Ⅱ. MATERIAL AND METHODS ·············································································· 3 A. Cells and cell culture ····························································································· 3 B. RT-PCR ················································································································· 3 C. Membrane preparation and western blotting ························································· 4 D. ERK/AKT Western blot analysis ·········································································· 4 E. Cell proliferation assays ························································································ 5 F. Neurosphere assay ································································································· 5 G. In vivo tumor growth ···························································································· 5 Ⅲ. RESULTS ················································································································· 7 A. HECAT5 overexpression increases neurosphere formation in cell type-specific manner ···················································································································· 7 B. Overexpression of HECAT5 increases tumorigenicity in cell type-specific manner ····························································································································· 11 C. Overexpression of HECAT5 promotes xenograft tumor formation ····················· 13 D. mRNA and protein expression levels of the muscarinic acetylcholine receptor 3 in each cell lines ····································································································· 15 E. Regulation of cell proliferation by M3R modulators ······································ 17 F. Cellular proliferation inhibited by M3R selective antagonists ····························· 19 G. Schematic indicating the phospho-acceptor sites in M3R and expression levels ······························································································································ 21 H. Differential expression of ERK and AKT pathways in three cell lines and principle of GPCR signaling ······························································································· 23 J. A different signaling pathways of M3R-HECAT5 in cell type-specific manner 25 Ⅳ. DISCUSSION ······································································································ 27 Ⅴ. CONCLUSION ···································································································· 29 REFERENCES ·········································································································· 30 국문요약 ····················································································································· 32 | - |
dc.language.iso | eng | - |
dc.publisher | The Graduate School, Ajou University | - |
dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
dc.title | 세포유형에 따른 HECAT5와 M3R의 상호작용을 통한 세포증식 조절 | - |
dc.title.alternative | HECAT5 regulates cell proliferation via interaction with M3R in cell type-specific manner | - |
dc.type | Thesis | - |
dc.contributor.affiliation | 아주대학교 일반대학원 | - |
dc.contributor.alternativeName | Woo Chang Sung | - |
dc.contributor.department | 일반대학원 의생명과학과 | - |
dc.date.awarded | 2016. 8 | - |
dc.description.degree | Master | - |
dc.identifier.localId | 758764 | - |
dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000022803 | - |
dc.subject.keyword | Embryonic stem cell | - |
dc.subject.keyword | Self-renewal | - |
dc.subject.keyword | Pluripotency | - |
dc.subject.keyword | Cell proliferation | - |
dc.subject.keyword | Stemness | - |
dc.description.alternativeAbstract | Embryonic stem (ES) cells are derived from pluripotent inner cell mass of the early mammalian embryo. Their self-renewal and pluripotency make them attractive sources for stem cell therapies. For ethical and practical reasons, ES cells have a major threat limiting in potential therapeutic applications. To investigate the mechanism regulating stemness, especially cell cycle of ES cells, our previous study identified HECAT5 which is co-expressed in both ES and cancer cells, but not normal tissues by using Digital Differential Display (DDD). Overexpression of HECAT5 promotes self-renewal capacity and it enhances tumorigenicity in cell type-specific manner. HECAT5 has effects on those through the interaction with M3R (Muscarinic acetylcholine receptor 3), candidate interacting partner of HECAT5, as modulators of M3R activities regulate cell proliferation in a HECAT5-dependent manner. In addition, overexpressed HECAT5 have different effects on the phosphorylation level of M3R in each cell types. In F3 cells, phosphorylation level of M3R was relatively low but HECAT5 enhanced its level. Meanwhile, in HEK293 cells, basically its level was high but HECAT5 decreased it. Besides, F3 and HEK293 cells showed different signaling pathways, ERK and AKT. Collectively, our study uncovers that a novel HECAT5 gene may play a role in regulating the stemness properties in cell type-specific manner. | - |
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