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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 廖憶純(Yi-Chun Liao) | |
| dc.contributor.author | Yen-kang Huang | en |
| dc.contributor.author | 黃彥康 | zh_TW |
| dc.date.accessioned | 2021-06-08T02:16:56Z | - |
| dc.date.copyright | 2015-12-01 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-10-16 | |
| dc.identifier.citation | 五、 參考文獻
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'Intracellular sorting of neuromodulin (GAP-43) mutants modified in the membrane targeting domain.' J Neurosci 14(10): 5807-5817. Lo, S. H. (2004). 'Tensin.' Int J Biochem Cell Biol 36(1): 31-34. Lo, S. H. and T. B. Lo (2002). 'Cten, a COOH-terminal tensin-like protein with prostate restricted expression, is down-regulated in prostate cancer.' Cancer Res 62(15): 4217-4221. Lo, S. H., Q. C. Yu, L. Degenstein, L. B. Chen and E. Fuchs (1997). 'Progressive kidney degeneration in mice lacking tensin.' J Cell Biol 136(6): 1349-1361. Owen, G. R., D. O. Meredith, I. ap Gwynn and R. G. Richards (2005). 'Focal adhesion quantification - a new assay of material biocompatibility? Review.' Eur Cell Mater 9: 85-96; discussion 85-96. Resh, M. D. (1994). 'Myristylation and palmitylation of Src family members: the fats of the matter.' Cell 76(3): 411-413. Resh, M. D. (1999). 'Fatty acylation of proteins: new insights into membrane targeting of myristoylated and palmitoylated proteins.' 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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/19749 | - |
| dc.description.abstract | 摘要
C-terminal tensin-like (CTEN) 為位於 focal adhesion 上tensin 家族的一員，除了前列腺和胎盤組織外，在其他正常組織中表現不多，但是在許多的癌症，如肺癌、大腸癌以及乳癌中卻有表現量上升的情況，除了 focal adhesion，CTEN在大腸癌細胞株 SW480 與 HCT 116 中有在核內累積的現象。在細胞核中大量存在的 CTEN 也被證實會和 Wnt pathway 中的 beta-catenin 結合並促進腫瘤發生。另一方面，若在正常的人類腎臟胚胎細胞 293A 與正常的前列腺細胞 RWPE-1 表現 CTEN，則會在細胞質內分布較多，顯示 CTEN 在細胞內的位置與細胞癌化的高度相關性。本論文假設 CTEN 在核內的累積會促使細胞癌化。藉由在 CTEN 蛋白質上加上 nucleus localization signal (NLS) 或 Src myristoylation signal，改變 CTEN 在細胞內的分布，並了解 CTEN 分布的差異對細胞癌化特性的影響。首先建立穩定表現帶有特殊胺基酸訊號 (NLS signal and Src myristoylation signal) 的 CTEN 蛋白質之細胞株，以西方墨點法 (western blot) 與免疫螢光法 (immunofluorescence) 確認在細胞內表現含不同訊號的CTEN 時，確實會導致 CTEN 在細胞內的分布有相對應的改變。接著以 cell proliferation assay 初步觀察不同 CTEN 分佈對細胞生理的影響，發現 CTEN 於細胞核內累積並不影響細胞生長速率。 | zh_TW |
| dc.description.abstract | C-terminal tensin-like (CTEN) locates in focal adhesion and belongs to tensin family. CTEN expresses much in prostate and placenta, but rarely expresses in other normal tissues. Elevated CTEN level has been detected in lung cancer, colorectal cancer and breast cancer. In addition to focal adhesion, CTEN also accumulates in the nucleus in colorectal cancer cell lines SW480 and HCT 116. It has been found that CTEN in nucleus interacts with Wnt pathway signal beta-catenin and promotes tumorigenesis. On the other hand, human kidney embryonic cell 293A and normal prostate cell RWPE-1 express CTEN predominantly in cytoplasm. It is shown that CTEN localization is related to tumorigenesis. We assumpted that CTEN accumulation in the nucleus will lead to tumor. By adding special signaling amino acid sequence, nucleus localization signal (NLS) and Src myristoylation signal cause CTEN to change localization in cell. Therefore, we can observe tumorigenesis of cells with different CTEN localization. First, we established stable cell lines expressing CTEN protein with special amino acid signals. We checked with western blot and immunofluorescence, and we confirmed that expressing CTEN with different signals does cause CTEN distribution changes. Then, we used cell proliferation assay to observe the effect of CTEN distribution in tumorigenesis, and we found that CTEN accumulation in cell nucleus do not affect cell proliferation rate. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T02:16:56Z (GMT). No. of bitstreams: 1 ntu-104-R02b22005-1.pdf: 2395478 bytes, checksum: 5a8e391bd409074453dfc252d90a5495 (MD5) Previous issue date: 2015 | en |
| dc.description.tableofcontents | 目錄
目錄 I 縮寫表 IV 摘要 VI Abstract VII 一、 研究背景 1 1.1 Focal adhesion 1 1.2 Tensin 2 1.3 CTEN 與癌症 2 1.4 蛋白質進出和與癌症的關係 3 1.5 協助蛋白質定位的特殊胺基酸序列 5 1.5.1 Membrane-targeting signaling sequence ──Src myristoylation 5 1.5.2 Nucleus-targeting signaling sequence ── Nuclear localization sequence (NLS) 6 1.6 研究目的 8 二、材料與方法 9 2.1 細胞培養 9 2.2 細胞繼代 9 2.3 細胞之冷凍與解凍 10 2.4 細胞計數 10 2.5 細胞轉染 11 2.6 表現 CTEN 的質體 DNA 建構與分析 11 2.6.1 聚合酶連鎖反應 (PCR) 11 2.6.2 DNA 片段純化 12 2.6.3 以限制酶截切質體與PCR產物 12 2.6.4 DNA 接合法 (DNA ligation) 12 2.6.5 重組質體之轉型與篩選 12 2.6.6 質體純化 13 2.6.7 以限制酶進行質體分析 13 2.6.8 DNA 瓊脂糖膠體電泳法 13 2.7 細胞全蛋白質之抽取 13 2.8 細胞質與細胞核萃取液分離 (fractionation) 14 2.9 蛋白質分析 14 2.9.1 蛋白質定量 14 2.9.2 聚丙烯醯胺膠體電泳 14 2.9.3 Coomassie Brilliant Blue (CBR) 染色法 15 2.9.4 蛋白質轉印 15 2.9.5 免疫呈色法 15 2.10 螢光染色 16 2.11 篩選穩定表現細胞株 16 2.12 Cell proliferation assay 17 三、研究結果 18 3.1 帶有特殊胺基酸訊號之 CTEN 在細胞內的分佈 18 3.2 重新建構帶有進核特殊胺基酸序列的 NLS-CTEN 20 3.3 建立穩定表現含不同訊息胺基酸序列 CTEN 的細胞株 20 3.4 確認 CTEN 在各組細胞株中的分佈 22 3.5 分析 CTEN 在細胞核的分佈是否影響細胞增生 23 四、討論與未來方向 24 五、參考文獻 28 六、圖與表 31 | |
| dc.language.iso | zh-TW | |
| dc.title | 細胞核中的 CTEN 於腫瘤發生過程中所扮演的角色 | zh_TW |
| dc.title | The Role of Nuclear CTEN in Tumorigenesis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃楓婷(Feng-Ting Huang),謝淑貞(Shu-Chen Hsieh),賴韻如(Yun-Ju Lai) | |
| dc.subject.keyword | 腫瘤發生,與細胞膜結合,進核,癌症,蛋白質分佈,細胞生長速率, | zh_TW |
| dc.subject.keyword | Tumorigenesis,CTEN,tensin,distribution,localization,cancer,tumor,membrane-targeting,nucleus-targeting,cell proliferation rate, | en |
| dc.relation.page | 55 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2015-10-16 | |
| 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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