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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 廖憶純(Yi-Chun Liao) | |
| dc.contributor.author | Chih-Hsuan Lin | en |
| dc.contributor.author | 林芷萱 | zh_TW |
| dc.date.accessioned | 2021-07-11T14:36:08Z | - |
| dc.date.available | 2022-09-04 | |
| dc.date.copyright | 2017-09-04 | |
| dc.date.issued | 2017 | |
| dc.date.submitted | 2017-08-18 | |
| dc.identifier.citation | 張佳鈞 (2013) 表皮生長因子促進 CTEN 基因表現之機制探討，碩士論文，國立臺灣大學生命科學院生化科技學系
劉益洲 (2015) p300 參與表皮生長因子促進 CTEN 基因表現之調控機制，碩士論文，國立臺灣大學生命科學院生化科技學系 潘妍卉 (2016) CTEN 磷酸化與其於核質間穿梭之調控機制，碩士論文，國立臺灣大學生命科學院生化科技學系 Albasri, A., Seth, R., Jackson, D., Benhasouna, A., Crook, S., Nateri, A. S., Chapman, R. and Ilyas, M. (2009). C-terminal Tensin-like (CTEN) is an oncogene which alters cell motility possibly through repression of E-cadherin in colorectal cancer. J Pathol 218(1): 57-65. Auger, K. R., Songyang, Z., Lo, S. H., Roberts, T. M. and Chen, L. B. (1996). Platelet-derived growth factor-induced formation of tensin and phosphoinositide 3-kinase complexes. J Biol Chem 271(38): 23452-23457. Barbieri, I., Pensa, S., Pannellini, T., Quaglino, E., Maritano, D., Demaria, M., Voster, A., Turkson, J., Cavallo, F., Watson, C. J., Provero, P., Musiani, P. and Poli, V. (2010). Constitutively active Stat3 enhances neu-mediated migration and metastasis in mammary tumors via upregulation of Cten. Cancer Res 70(6): 2558-2567. Bennett, D. T., Reece, T. B., Foley, L. 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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77857 | - |
| dc.description.abstract | 表皮生長因子受體 (EGFR) 訊息傳導的活化能促進 CTEN 基因表達，並增強乳癌細胞的遷移能力與侵犯性，但其調控 CTEN 基因表達之機制目前尚未清楚。我們先前的研究證實，在人類子宮頸癌細胞株 HeLa 中，EGFR的活化會透過 MEK/ERK訊息傳導路徑，藉由組蛋白乙醯轉移酶p300調控 CTEN 啟動子上組蛋白的乙醯化，並促進 p300 與 CTEN 啟動子的結合，進而誘導 CTEN 的表現。本論文進一步研究發現，將 p300 受 ERK2 磷酸化之胺基酸位點突變，會降低 p300 與 CTEN 啟動子的結合，並減少 EGF 誘導 CTEN 之 mRNA及蛋白質表現量，證實 EGFR 訊息傳導活化，經由調控 p300 的磷酸化而影響與 CTEN 啟動子的結合，進而促進 CTEN 基因表達。後續針對與 p300 有結合能力且在 CTEN 啟動子上有預測結合位之轉錄因子 Sp1 進行深入探討，結果證實 HeLa 細胞中 p300 及 Sp1 確實有交互作用，且 EGF 能增進 Sp1 與 CTEN 啟動子的結合。另一方面，也證實 EGFR 能透過 MEK/ERK 及 PI3K 訊息傳導路徑，誘導 CTEN 蛋白質 Thr347、Ser350、Ser386 位點之磷酸化，初步結果發現 CTEN 磷酸化與否並不影響 HeLa 細胞之增生能力，磷酸化的CTEN 與促進腫瘤發生的相關性有待更深入的探討。 | zh_TW |
| dc.description.abstract | CTEN gene expression is upregulated by the activation of epidermal growth factor receptor (EGFR), and CTEN promotes cell migration and invasion in breast cancer. However, the mechanism of EGFR-induced CTEN gene expression is still unclear. On the basis of our previous studies, we have demonstrated that EGFR activation induces histone acetylation on CTEN promoter by histone acetyltransferase p300 through MEK/ERK signaling pathway in the human cervical cancer cell line, HeLa. EGF stimulation also recruits p300 binding to CTEN promoter and results in up-regulation of CTEN expression. In this study, p300 with mutations on the amino acid residues that can be phosphorylated by ERK2 was constructed. We discovered that recruitment of the mutant p300 toward CTEN promoter ia decreased and the levels of CTEN mRNA and proteininduced by EGF are reduced in the presence of the mutant p300. It indicates that EGFR signaling can regulate the recruitment of p300 toward CTEN promoter and further activate CTEN gene expression through phosphorylating p300. In addition, we investigated whether the Sp1 transcription factor, which has putative binding site on CTEN promoter, can interact with p300 and bind to CTEN promoter. Our results have shown that after EGF stimulation, the recruitment of Sp1 to CTEN promoter is enhanced. Furthermore, we demonstrated the interaction between p300 and Sp1 in HeLa cell. On the other hand, we confirmed that EGFR can induce the phosphorylation of T347, S350, S386 sites on CTEN protein via MEK/ERK pathway and PI3K pathway. Our preliminary result has shown that CTEN phosphorylation has no effect on the proliferation of HeLa cell. However, the correlation between CTEN phosphorylation and tumorigenesis warrants further investigation. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T14:36:08Z (GMT). No. of bitstreams: 1 ntu-106-R04B22016-1.pdf: 1713048 bytes, checksum: 283f134b0465ada35a0d18a1cd3b3ed2 (MD5) Previous issue date: 2017 | en |
| dc.description.tableofcontents | 縮寫表 iii
摘要 v Abstract vi 1 本論文之研究基礎 1 1.1 表皮生長因子受體訊息傳導路徑 1 1.2 Tensin 家族 2 1.3 C-terminal tensin like (CTEN) 蛋白質功能研究 3 1.4 EGFR 訊息傳導路徑調控CTEN 基因表達之機制 4 1.5 EGFR 訊息傳導路徑調控 CTEN 蛋白質磷酸化 5 1.6 E1A binding protein p300 蛋白質功能研究 6 1.7 轉錄因子 Sp1 蛋白質功能研究 7 1.8 本論文研究目的 8 2 實驗材料與方法 9 2.1 實驗材料 9 2.1.1 菌種 9 2.1.2 質體DNA 9 2.1.3 細胞株 11 2.1.4 細胞培養基 11 2.1.5 表皮生長因子及激酶抑制劑 11 2.2 實驗方法 12 2.2.1 細胞相關實驗 12 2.2.2 DNA 分析及質體建構 15 2.2.3 RNA 分析 17 2.2.4 蛋白質分析 17 2.2.5 轉錄因子預測結合位線上分析 19 2.2.6 染色質免疫沉澱 (Chromatin Immunoprecipitation, ChIP) 20 2.2.7 免疫共沉澱 (Co-Immunoprecipitation, Co-IP) 22 3 研究結果 23 3.1 EGFR 訊息傳導的活化可使 p300 磷酸化進而增加 p300 與 CTEN 啟動子的結合 23 3.2 EGFR 訊息傳導的活化會藉由 p300 的磷酸化促進 CTEN 基因表達 25 3.3 EGFR 訊息傳導的活化 促進 Sp1 與CTEN 啟動子結合 25 3.4 利用免疫共沉澱法證實 p300 與 Sp1 在 HeLa 細胞中有交互作用 26 3.5 EGFR 訊息傳導的活化可誘導 CTEN 磷酸化 27 3.7 調控CTEN 磷酸化之訊息傳導路徑 30 3.8 CTEN 磷酸化對細胞增殖的影響 31 4 討論與未來研究方向 32 5 參考文獻 36 6 圖與表 48 | |
| dc.language.iso | zh-TW | |
| dc.subject | 磷酸化 | zh_TW |
| dc.subject | 轉錄因子 | zh_TW |
| dc.subject | 表皮生長因子 | zh_TW |
| dc.subject | 基因調控 | zh_TW |
| dc.subject | CTEN | en |
| dc.subject | EGF | en |
| dc.subject | p300 | en |
| dc.subject | Sp1 | en |
| dc.subject | phosphorylation | en |
| dc.title | 表皮生長因子促進 CTEN 基因表達及磷酸化之調控機制 | zh_TW |
| dc.title | Study of the regulatory mechanism of EGF-induced upregulation and phosphorylation of CTEN | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張麗冠(Li-Kwan Chang),謝淑貞(Shu-Chen Hsieh),黃楓婷(Feng-Ting Huang) | |
| dc.subject.keyword | 基因調控,表皮生長因子,轉錄因子,磷酸化, | zh_TW |
| dc.subject.keyword | CTEN,EGF,p300,Sp1,phosphorylation, | en |
| dc.relation.page | 66 | |
| dc.identifier.doi | 10.6342/NTU201703908 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2017-08-19 | |
| 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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