請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77846完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 廖憶純 | |
| dc.contributor.author | Yi-Xuan Wang | en |
| dc.contributor.author | 王乙喧 | zh_TW |
| dc.date.accessioned | 2021-07-11T14:35:55Z | - |
| 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 | 李昌恆 (2014) 探討 Cten 與 β-catenin 和 α-actinin4 之間的交互作用及 Cten 於細胞質和細胞核間穿梭的機制，碩士論文，國立台灣大學生命科學院生化科技學系
莊惟 (2015) CTEN 於細胞核中的功能探討，碩士論文，國立台灣大學生命科學院生化科技學系 黃彥康 (2015) 細胞核中的 CTEN 於腫瘤發生過程中所扮演的角色，碩士論文，國立台灣大學生命科學院生化科技學系 邱品慈 (2016) 探討細胞核中 CTEN 參與調控 NF-κB 及 ERα 訊號反應路徑之功能，碩士論文，國立台灣大學生命科學院生化科技學系 Al-Ghamdi, S., Cachat, J., Albasri, A., Ahmed, M., Jackson, D., Zaitoun, A., Ilyas, M. (2013). C-terminal tensin-like gene functions as an oncogene and promotes cell motility in pancreatic cancer. Pancreas, 42(1), 135-140. A-Ri, C., Uchio-Yamada, K., Takeshi, T., Tomomi, M., Ichiro, M., Junichiro, M., Tsutomu, K.,Yasuhiro, K., Atsushi, A., Nobuya, S., Takashi, A. (2006). Deficiency of the tensin2 gene in the ICGN mouse: an animal model for congenital nephrotic syndrome. Mammalian Genome, 17(5), 407-416. Archbold, H. C., Yang, Y. X., Chen, L. and Cadigan, K. M. (2012). How do they do Wnt they do? : regulation of transcription by the Wnt/beta-catenin pathway. Acta Physiol (Oxf), 204(1), 74-109. Cance, W. G., Harris, J. E., Iacocca, M. V., Roche, E., Yang, X., Chang, J., Xu, L. (2000). Immunohistochemical analyses of focal adhesion kinase expression in begin and malignant human breast and colon tissue: correlation with preinvasive and invasive phenotypes. Clin Cancer Res, 6(6), 2417-2423. Calderwood, D. A., Fujioka, Y., De Pereda, J. M., Garcia-Alvarez, B., Nakamoto, T., Margolis, B. and Ginsberg, M. H. (2003). Integrin beta cytoplasmic domain interactions with phosphotyrosine-binding domains: a structure prototype for diversity in integrin signaling. Proc Natl Acad Sci U S A, 100(5), 2272-2277. Chen, H., Duncan, I. C., Bozorgchami, H. and Lo, S. H. (2002).Tensin1 and a previously undocumented family membrane, tensin2, positively regulate cell migration. Proc Natl Acad Sci U S A, 99(2), 733-738. Christian, D., Bouatia-Naji, N., Nathan, T., Francesca, N. D., Katelynn, T., Ronen, D., Maelle, P., Fernandez-Friera, L., Jorge, S., Thierry, L. T., Ming-Huei, C., Vincent, P., Yohan, B., Philippe, P., Diana, Z., Mark, L., Serge, H., Ronan, R., Emelia, J. B., Fabrice, B., Su, H. Lo., Elena, D., Floriane, S., Simon, L. (2015). Genetic association analyses highlight biological pathways underlying mitral valve prolapse. Nature Genetics, 47, 1206-1211. Chiang, M. K., Liao, Y. C., Kuwabara, Y. and Lo S. H. (2005). Inactivation of tensin3 in mice results in growth retardation and postnatal lethality. Dev Biol, 279(2) 368-377. Clevers, H. (2006). Wnt/beta-catenin signaling in development and disease. Cell, 127(3), 469-480. Davis, S., Lu, M. L., Lo, S. H., Lin, S., Butler, J. A. Druker, B. J. and Chen, L. B. (1991). Presence of an SH2 domain in the actin-binding protein tensin. Science, 252(5006), 712-715. Eli, Z. and Benjamin, G. (2001). Molecular complexity and dynamics of cell-matrix adhesions. J Cell Science, 114, 3583-3590. Gorlich, D. and Ulrike, K. (1999). Transport between the cell nucleus and the cytoplasm. Annu Rev Cell Dev Biol, 15, 607-660. Ishii, A. and Lo S. H. (2001). A role of tensin in skeletal-muscle regeneration. Biochem J, 356(3), 737-745. Kassel, O., Schneider, S., Heilbock, C., Litfin, M., Gottlicher, M., Herrlich, P., (2004). A nuclear isoform of the focal adhesion LIM-domain protein Trip6 integrates activating and repressing signals at AP-1- and NF-kappaB-regulated promoters. Genes Dev, 18(20),2518-2528. Katz, M., Amit, I., Citri, A., Shay, T., Carvalho, S., Lavi, S. and Yarden, Y. (2007). A reciprocal tensin-3-cten switch mediates EGF-driven mammary cell migration. Nat Cell Biol, 9(8), 961-969. Ke R., Xiao L., Nan Y., Yong C., Aizhen Y., Hui C., Jian Z., Sujia W., Xin S., Chen W., Xiaoliang S. (2015). Focal adhesion kinase overexpression and its impact on human osteosarcoma. Oncotarget, 6, 31085-31103. Kustatscher, G., Wills, K. L., Furlan, C., Rappsilber, J. (2014). Chromatin enrichment for proteomics. Nat Protoc, 9(9), 2090-2099. la Cour, T., Kiemer, L., Molgaard, A., Gupta, R., Skriver, K., Brunak, S., (2004). Analysis and prediction of leucine-rich nuclear export signals. Protein Eng Des, 17, 527-536. Li, C., M. Iida, E. F. Dunn, A. J. Ghia and D. L. Wheeler (2009). Nuclear EGFR contributes to acquired resistance to cetuximab. Oncogene, 28(43): 3801-3813. Liao, Y. C., Chen, N. T., Shih, Y. P., Dong, Y. and Lo, S. H. (2009). Up-regulation of C-terminal tensin-like molecule promotes the tumorigenicity of colon cancer through beta-catenin. Cancer Res, 69(11), 4563-4566. Lo and Lo, T. B. (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. (2004). Tensin. Int J Biochem Cell Biol, 36(1), 31-34. Lo, S. H., Janmey, P. A., Hartwig, J. H. and Chen, L. B. (1994). Interaction of tensin with actin and identification of its three distinct actin-binding domains. J Cell Biol, 125(5), 1067-1075. Lo, S. H., Yu, Q. C., Degenstein L., Chen, L. B. and Fuchs, E. (1997). Progressive Kidney Degeneration in Mice Lacking Tensin. J Cell Biol, 136(6), 1349-1316. Makkerh, J. P., Dingwall, C., Laskey, R.A. (1996). Comparative mutagenesis of nuclear localisation signals reveals the importance of neutral and acidic amino acids. Curr Biol, 6(8), 1025–1027. Maurer-Stroh, S., Eisenhaber, B., Eisenhaber, F. (2002). N-terminal N-myristoylation of proteins: prediction of substrate proteins from amino acid sequence. J Mol Biol, 317(4), 541-57. Siesser, P. M., Hanks, S. K., (2006). The signaling and biological implications of FAK overexpression in cancer. Clin Cancer Res, 12, 3233-3237. Rajfur, Z., Roy, P., Otey, C., Romer, L. and Jacobson, K. (2002). Dissecting the link between stress fibers and focal adhesions by CALI with EGFP fusion proteins. Nat cell Biol, 4(4), 286-293. Resh, M. D. (1994). Myristylation and palmitylation of Src family members: The fats of the matter. Cell, 76, 411-413. Resh, M. D. (1999). Fatty acylation of proteins: new insights into membrane targeting of myristoylated and palmitoylated proteins. Biochim Biophys Acta, 1451(1), 1-16. Sakashita, K., Moriyma, K., Tanaka, F., Kamohara, Y., Inoue., Sawada, T., Mori, M. (2008). Prognostic relevance of Tensin4 expression in human gastric cancer. Ann Surg Oncol, 15(9), 2606-2613. Sasaki, H., Moriyama, S., Mizuno, K., Yukiue, H., Konishi, A., Yano, M., Fujii, Y. (2003). Cten mRNA expression was correlated with tumor progression in lung cancers. Lung cancer, 40(2), 151-155. Sasaki, H., Yukiue, H., Kobayashi, Y., Fukai, I. and Fujii, Y. (2003). Cten mRNA expression is correlated with tumor progression in thymoma. Tumor Biol, 24(5), 271-274. Satoko, Y., Tsutomu M., Shuhei Y., Naohito U., Seishiro W., Hisashi U., Hitoshi Y., Tsuyoshi M., Kazuo E., Toshiaki N., Kazutaka K., Shigeki K. (2005). Enhanced expression of p46 Shc in the nucleus and p52 Shc in the cytoplasm of human gastric cancer. Int J Oncol, 26(4), 905-911. Stewart, M. (2007). Molecular mechanism of the nuclear protein import cycle. Nat Rev Mol Cell Biol, 8, 195-208. Timothy, J. Y. (2004). A renaissance for SRC. Nature Rev Cancer, 4, 470-480 Werneburg, B. G., Zoog, S. J., Dang, T. T. A., Kehry, M. R. and Crute, J. J. (2001). Molecular Characterization of CD40 Signaling Intermediates. J. Biol. Chem, 276, 43334-43342. Xiaolan, Z., Man, C., Jishan, T., (2016). Knockdown of ZFR suppresses cell proliferation and invasion of human pancreatic cancer. Biol Res, 49, 26. Yang, K., Wu W. M., Chen, Y.C, Lo, S. H., Liao, Y. C. (2016). ΔNp63α Transcriptionally Regulates the Expression of CTEN That Is Associated with Prostate Cell Adhesion. PLoS One, 11(1). Zaidel-Bar, R., Cohen M., Addadi L. and Geiger, B. (2004). Hierarchical assembly of cell-matrix adhesion complexes. Biochem Soc Trans, 32, 416-420. Zhongping, Gu., Liran, Z., Shen, G., Zhengxin, W. (2001). Nuclear Transport Signals Control Cellular Localization and Function of Androgen Receptor Cofactor p44/WDR77. PLoS One, 6(7). | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77846 | - |
| dc.description.abstract | C-terminal tensin-like (CTEN) 蛋白質為 tensin 家族的成員之一，位在 focal adhesion 上，參與細胞的貼附、增生、遷移等行為，在許多癌症中皆發現 CTEN 高度表現。根據我們之前的研究顯示，相較於正常細胞中 CTEN 主要位在細胞質及 focal adhesion，大量的 CTEN 卻被發現聚集在癌細胞的細胞核中，顯示 CTEN 在細胞中的定位與腫瘤的發生具有高度的相關性。本論文假設 CTEN 在細胞核中累積會促進腫瘤發生，藉由在 CTEN 蛋白質加上幫助定位的胺基酸序列 nucleus localization signal (NLS) 或 Src myristoylation signal (myr)，進而改變 CTEN 在細胞內的分布，以了解 CTEN 分布的差異對細胞腫瘤形成特性的影響。實驗結果發現，在沒有內生性 CTEN 的 293A 細胞株中，穩定表現不同細胞定位的外源性 CTEN，細胞的生長與貼附能力並沒有因此而產生差異。而在只有少量內生性 CTEN 的大腸癌細胞株 HCT116 中，穩定表現於細胞核內累積的外源性 CTEN 不會影響 HCT116 細胞生長速率，但是會增加細胞移動與入侵能力。另一方面，我們先前發現在細胞核中的 CTEN 可能會與 DNA 結合，因此使用表現大量內生性 CTEN 的大腸癌細胞 HT29，以 chromatin enrichment proteomics (ChEP) assay 分析確認細胞核中的 CTEN 的確會與 DNA 結合，最後進行 chromatin immunoprecipitation sequencing (ChIP-seq) 找出 CTEN 結合的目標 DNA。 | zh_TW |
| dc.description.abstract | C-terminal tensin-like (CTEN) protein locates at focal adhesion and belongs to tensin family. It involve in cell adhesion, proliferation and migration. Elevated CTEN level has been detected in many cancers. On the basis of our previous studies, a high population of CTEN accumulates in the nucleus of cancer cells whereas it is predominantly associated with focal adhesions and localized in cytoplasm in normal cells. It suggests that the subcellular localization of CTEN is highly related to tumorigenesis. In this study, we hypothesize that the nuclear accumulation of CTEN contributes to tumorigenesis. By adding subcellular targeting sequences, such as nucleus localization signal (NLS) or Src myristoylation signal (myr), we manipulate the localization of CTEN to study the effect of different subcellular distributions of CTEN on cell tumorigenicity. The stably-expressed modified CTEN variants in 293A cells, which have no endogenous CTEN, did not affect cell adhesion and proliferation. However, when the stably-expressed CTEN accumulated in the nucleus in HCT116 colon cancer cell line, the cell migration and invasion ability were increased but the proliferation rate was not affected. On the other hand, our previous work found that nuclear CTEN may bind to DNA. Therefore, chromatin enrichment proteomics (ChEP) assay was first applied in HT29 colon cancer cell line to confirm the interaction of CTEN and DNA. Finally, chromatin immune-precipitation sequencing (ChIP-seq) was used to identify the target DNAs that bind to CTEN. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T14:35:55Z (GMT). No. of bitstreams: 1 ntu-106-R04b22054-1.pdf: 2632790 bytes, checksum: 8cf244e3854e74a2678fe6350417cef7 (MD5) Previous issue date: 2017 | en |
| dc.description.tableofcontents | 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 family 2 1.3 C-terminal tensin like protein (CTEN) 蛋白質之功能研究 3 1.4 CTEN 蛋白質進出細胞核與癌症的相關性 4 1.5 CTEN 蛋白質於核質間穿梭的分子機制 4 1.6 研究目的 6 二、 材料與方法 8 2.1 Cell preparation 8 2.1.2 Cell lines 8 2.1.2 Cell culture medium 8 2.1.3 Cell culture 8 2.1.4 Cell subculture 9 2.1.5 Freezing and Thawing 9 2.1.6 Cell counting 10 2.2 Plasmids and Transfection 10 2.2.1 CTEN expression plasmids 10 2.2.2 Transfection 11 2.3 Stable cell selection and assays 11 2.3.1 Stable cell line selection 12 2.3.2 Cell proliferation assay 13 2.3.3 Cell adhesion assay 13 2.3.4 Cell migration assay 13 2.3.5 Cell invasion assay 14 2.4 Subcellular fractionation 15 2.5 Immunofluorescence staining 15 2.6 Protein analysis 16 2.6.1 SDS PAGE 16 2.6.2 Western blotting 17 2.6.3 Immunoblotting 17 2.7 Chromatin enrichment for proteomics assay (ChEP assay) 17 2.7.1 Formaldehyde cross-linking and cell harvesting 17 2.7.2 Cell lysis and RNase digestion 18 2.7.3 Chromatin enrichment 18 2.8 Chromatin immunoprecipitation sequencing (ChIP seq.) 19 2.8.1 Crosslinking and cell pellet isolation 19 2.8.2 Lysis and microccal nuclease (Mnase) digestion 19 2.8.3 Protein/DNA immune-precipitation 20 2.8.4 IP elution 20 2.8.5 DNA recovery 20 2.8.6 DNA library construction 20 2.8.7 NGS 20 2.8.8 CLC 軟體分析實驗結果 21 三、 研究結果 22 3.1 WT-CTEN, myr-CTEN 與 NLS-CTEN 在 293A 細胞內的分布 22 3.2 分析 CTEN 在細胞內的分布是否影響 293A 細胞株增生與貼附能力 23 3.3 WT-CTEN, myr-CTEN 與 NLS-CTEN在 RWPE-1 細胞內的分布 24 3.4 WT-CTEN, myr-CTEN 與 NLS-CTEN 在 HCT116 細胞內的分布 25 3.5 分析 CTEN 在細胞內的分布是否影響 HCT116 細胞株增生、移動與 侵襲能力 27 3.6 WT-CTEN, myr-CTEN 與 NLS-CTEN在 DLD-1 細胞內的分布 27 3.7 分析 CTEN 在細胞內的分布是否影響 DLD-1 細胞株增生、移動 能力 29 3.8 利用 ChEP assay 探討在CTEN 蛋白質是否結合於 DNA 上 29 3.9 利用 ChIP Sequence 分析 CTEN 與 DNA 的結合序列 31 3.9.1 最適 Chromatin fragmentation 條件探討 31 3.9.2 以毛細管電泳分析 ChIP DNA 32 3.9.3 NGS 定序分析 33 四、 討論與未來方向 35 五、 參考資料 42 六、 圖與表 47 附圖 77 | |
| dc.language.iso | zh-TW | |
| dc.subject | DNA 結合 | zh_TW |
| dc.subject | 進核 | zh_TW |
| dc.subject | 腫瘤發生 | zh_TW |
| dc.subject | 蛋白質分布 | zh_TW |
| dc.subject | 細胞移動及入侵能力 | zh_TW |
| dc.subject | Tumorigenesis | en |
| dc.subject | CTEN | en |
| dc.subject | tensin | en |
| dc.subject | distribution | en |
| dc.subject | localization | en |
| dc.subject | cancer | en |
| dc.subject | tumor | en |
| dc.subject | nucleus-targeting | en |
| dc.subject | cell migration and invasion ability | en |
| dc.subject | bind to DNA | en |
| dc.title | CTEN 於細胞核中累積對腫瘤形成特性的影響 | zh_TW |
| dc.title | Effect of the nuclear accumulation of CTEN on cell tumorigenicity | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張麗冠,謝淑貞,黃楓婷 | |
| dc.subject.keyword | 腫瘤發生,進核,蛋白質分布,細胞移動及入侵能力,DNA 結合, | zh_TW |
| dc.subject.keyword | Tumorigenesis,CTEN,tensin,distribution,localization,cancer,tumor,nucleus-targeting,cell migration and invasion ability,bind to DNA, | en |
| dc.relation.page | 87 | |
| dc.identifier.doi | 10.6342/NTU201703970 | |
| 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 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-106-R04b22054-1.pdf 未授權公開取用 | 2.57 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。