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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 廖憶純(Yi-Chun Liao) | |
dc.contributor.author | Ping-Tzu Chiu | en |
dc.contributor.author | 邱品慈 | zh_TW |
dc.date.accessioned | 2021-06-08T01:44:50Z | - |
dc.date.copyright | 2016-11-02 | |
dc.date.issued | 2016 | |
dc.date.submitted | 2016-08-15 | |
dc.identifier.citation | 李昌恆 (2014) 探討 CTEN 與 β-catenin 和 α-actinin4之間的交互作用及 CTEN 於細胞核質間穿梭的機制,碩士論文,國立台灣大學生命科學院生化科技學系
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/19093 | - |
dc.description.abstract | C-terminal tensin like (CTEN) 蛋白質,為 tensin 家族中的一員,位於 focal adhesion 上,參與細胞的貼附、增生、遷移等行為。 CTEN 在各個時期的大腸癌腫瘤組織中,皆有過量表現的現象,且於細胞核中也能偵測到高表現量的 CTEN。在人類大腸癌細胞株 SW480 中,CTEN 與 α-actinin4 於細胞核中有交互作用。過去文獻中指出,α-actinin4 在細胞核中作為 transcription factor co-activator 促進 nuclear factor kappa B (NF-κB) 及 estrogen receptor α (ERα) 的轉錄活性,影響其下游基因表現。因此,本研究探討 CTEN 是否參與調控 NF-κB 與 ERα 的訊息傳遞,藉此瞭解細胞核中 CTEN 的功能。本論文發現,在 SW480 中 knockdown CTEN 會導致 NF-κB 的轉錄活性下降,也抑制了下游目標基因 TRAF-1 及 IL-1β 之表現, 推測可能影響 SW480 大腸癌細胞之細胞凋亡及免疫反應。我們也證實 CTEN 不影響 NF-κB 的磷酸化與其在細胞核中的累積量,但與其 subunit, p65 在細胞核中有交互作用。另一方面,本論文證實,在 MCF-7 中過量表現 CTEN 會導致 ERα 的轉錄活性降低,也抑制了下游目標基因 PR、pS2 之表現,進而影響 MCF-7 乳癌細胞之細胞增生能力。 | zh_TW |
dc.description.abstract | C-terminal tensin like (CTEN) protein, a member of tensin family, locates at focal adhesion and participates in regulation of cell adhesion, proliferation and migration. Elevated CTEN level has been detected in all stage of colon cancers. Furthermore, a high population of CTEN is located in the nucleus and interacts with α-actinin4 in colon cancer cell line, SW480. Previous studies indicated that α-actinin4 might function as a transcription factor co-activator in nuclear factor kappa B (NF-κB) and estrogen receptor α (ERα) signaling pathway in the nucleus. In this study, we investigated that whether nuclear CTEN participates in NF-κB and ERα signaling pathway to elucidate the functional role of CTEN in the nucleus. Our results have shown that knockdown of CTEN down-regulates NF-κB reporter activity and decreases the expression of TRAF-1 and IL-1β, which are NF-κB signaling pathway downstream genes. It suggests that CTEN might affect cell apoptosis and immunity in SW480 colon cancer cells. However, CTEN has no effect on TNFα-induced phosphorylation of p65, IKKβ and IκBα after TNFα stimulation. We demonstrated that CTEN does not affect p65 nuclear translocation but interacts with p65 in the nucleus. Moreover, we found that CTEN repress ERα transcriptional activity. Overexpression of CTEN decreases the expression of ERα downstream genes, PR and pS2 and inhibits cell proliferation activity in MCF-7 breast cancer cells. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T01:44:50Z (GMT). No. of bitstreams: 1 ntu-105-R03b22046-1.pdf: 3531573 bytes, checksum: 3d66bc0cef4976e54816a155c97a46af (MD5) Previous issue date: 2016 | en |
dc.description.tableofcontents | 目錄 i
縮寫表 iv 摘要 vii Abstract viii 一、 本論文之研究基礎 1 1.1 Focal adhesion 1 1.2 Tensin Family 1 1.3 C-terminal tensin like protein (CTEN) 蛋白質之功能研究 2 1.4 α-actinin4 (ACTN4) 蛋白質之功能研究 4 1.5 Nuclear factor kappa B (NF-κB) 參與調控之訊息傳遞路徑 5 1.6 Estrogen receptor α (ERα) 參與調控之訊號反應路徑 6 1.7 本論文之研究目的 8 二、 材料與方法 9 2.1 菌種 9 2.2 質體 DNA 9 2.3 細胞相關實驗 10 2.4 DNA 分析及質體建構 13 2.5 RNA 分析 15 2.6 蛋白質分析 16 2.7 Dual luciferase assay 17 2.8 全細胞之蛋白質萃取 17 2.9 細胞核細胞質分離 (Fractionation) 17 2.10 免疫沉澱與免疫共沉澱 (Immunoprecipitation, IP and Co-IP) 18 2.11 線上資料庫分析 18 三、 研究結果 19 3.1 CTEN 參與調控 NF-κB 訊號反應路徑 19 3.1.1 利用Dual luciferase assay 找出活化 NF-κB 轉錄活性之最佳條件 19 3.1.2 CTEN 與 α-actinin4 參與調控 NF-κB訊號反應路徑 20 3.2 CTEN 與 α-actinin4 對於 NF-κB訊號反應路徑中下游目標基因表現的影響 20 3.2.1 CTEN 與 α-actinin4 藉由調控 NF-κB訊號反應路徑對細胞凋亡方面之影響 20 3.2.2 CTEN 與 α-actinin4 藉由調控 NF-κB訊號反應路徑對發炎及免疫方面之影響 22 3.3 CTEN 與α-actinin4 對於 NF-κB 相關蛋白質複合體的影響 22 3.3.1 CTEN 與 α-actinin4 對於 NF-κB訊號反應路徑中相關蛋白磷酸化之影響 22 3.3.2 CTEN 並不影響 p65 在細胞中的分布 23 3.3.3 利用免疫共沉澱法證實 CTEN 與 p65 於細胞核中有交互作用 24 3.4 CTEN 參與調控 ERα 訊號反應路徑 25 3.4.1 利用 Dual luciferase assay 找出活化 ERα 轉錄活性之最佳條件 25 3.4.2 證實 α-actinin4 參與調控 ERα 訊號反應路徑 25 3.4.3 CTEN 參與調控 ERα 訊號反應路徑,並抑制 ERα 之轉錄活性 26 3.5 乳癌細胞中 CTEN 參與調控 ERα 訊號反應路徑 27 3.5.1 在 MCF-7 細胞株中利用 Dual luciferase assay 找出活化 ERα 轉錄活性之最佳條件 27 3.5.2 在 MCF-7 細胞株中 CTEN 對於調控 ERα 訊號反應之影響 27 3.5.3 Kaplan-Meier Plotter 線上資料庫分析 CTEN 的表現與ERα-positive 乳癌病患存活率之相關性 28 3.6 CTEN 對於 ERα 訊號反應中下游目標基因表現的影響 28 3.6.1 CTEN 藉由調控 ERα 訊號反應路徑對細胞增殖的影響 28 3.6.2 在 MCF-7 細胞株中觀察 CTEN 對於調控細胞增生能力之影響 29 四、 討論與未來研究方向 30 五、 參考文獻 36 六、 圖與表 42 | |
dc.language.iso | zh-TW | |
dc.title | 探討細胞核中 CTEN 參與調控 NF-κB 及 ERα 訊號反應路徑之功能 | zh_TW |
dc.title | Functional relevance of nuclear CTEN in NF-κB and ERα signaling | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張麗冠(Li-Kwan Chang),黃楓婷(Feng-Ting Huang),謝淑貞(Shu-Chen Hsieh) | |
dc.subject.keyword | 訊號反應路徑,蛋白質交互作用,細胞凋亡,細胞增生, | zh_TW |
dc.subject.keyword | C-terminal tensin-like protein (CTEN),Nuclear-factor-kappa B (NF-κB),Estrogen receptor alpha (ERα), | en |
dc.relation.page | 84 | |
dc.identifier.doi | 10.6342/NTU201602533 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2016-08-16 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
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