請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23196完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 沈湯龍 | |
| dc.contributor.author | Lin-Ya Huang | en |
| dc.contributor.author | 黃麟雅 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:46:54Z | - |
| dc.date.copyright | 2009-08-03 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-07-30 | |
| dc.identifier.citation | Arner, E.S., and Holmgren, A. (2000). Physiological functions of thioredoxin and thioredoxin reductase. Eur J Biochem 267, 6102-6109.
Beckler, A., Moskaluk, C.A., Zaika, A., Hampton, G.M., Powell, S.M., Frierson, H.F., Jr., and El-Rifai, W. (2003). Overexpression of the 32-kilodalton dopamine and cyclic adenosine 3',5'-monophosphate-regulated phosphoprotein in common adenocarcinomas. Cancer 98, 1547-1551. Bertini, R., Howard, O.M., Dong, H.F., Oppenheim, J.J., Bizzarri, C., Sergi, R., Caselli, G., Pagliei, S., Romines, B., Wilshire, J.A., et al. (1999). Thioredoxin, a redox enzyme released in infection and inflammation, is a unique chemoattractant for neutrophils, monocytes, and T cells. J Exp Med 189, 1783-1789. Bieche, I., Tomasetto, C., Regnier, C.H., Moog-Lutz, C., Rio, M.C., and Lidereau, R. (1996). Two distinct amplified regions at 17q11-q21 involved in human primary breast cancer. Cancer Res 56, 3886-3890. Bown, N., Lastowska, M., Cotterill, S., O'Neill, S., Ellershaw, C., Roberts, P., Lewis, I., and Pearson, A.D. (2001). 17q gain in neuroblastoma predicts adverse clinical outcome. U.K. Cancer Cytogenetics Group and the U.K. Children's Cancer Study Group. Med Pediatr Oncol 36, 14-19. Dasgupta, S., Wasson, L.M., Rauniyar, N., Prokai, L., Borejdo, J., and Vishwanatha, J.K. (2009). Novel gene C17orf37 in 17q12 amplicon promotes migration and invasion of prostate cancer cells. Oncogene. Dean-Colomb, W., and Esteva, F.J. (2008). Her2-positive breast cancer: herceptin and beyond. Eur J Cancer 44, 2806-2812. Dieras, V., Vincent-Salomon, A., Degeorges, A., Beuzeboc, P., Mignot, L., and de Cremoux, P. (2007). [Trastuzumab (Herceptin) and breast cancer: mechanisms of resistance]. Bull Cancer 94, 259-266. Eklund, H., Gleason, F.K., and Holmgren, A. (1991). Structural and functional relations among thioredoxins of different species. Proteins 11, 13-28. Evans, E.E., Henn, A.D., Jonason, A., Paris, M.J., Schiffhauer, L.M., Borrello, M.A., Smith, E.S., Sahasrabudhe, D.M., and Zauderer, M. (2006). C35 (C17orf37) is a novel tumor biomarker abundantly expressed in breast cancer. Mol Cancer Ther 5, 2919-2930. Greenfield, N.J. (2006). Using circular dichroism spectra to estimate protein secondary structure. Nat Protoc 1, 2876-2890. Hayashi, T., Ueno, Y., and Okamoto, T. (1993). Oxidoreductive regulation of nuclear factor kappa B. Involvement of a cellular reducing catalyst thioredoxin. J Biol Chem 268, 11380-11388. Henson, E.S., Hu, X., and Gibson, S.B. (2006). Herceptin sensitizes ErbB2-overexpressing cells to apoptosis by reducing antiapoptotic Mcl-1 expression. Clin Cancer Res 12, 845-853. Holmgren, A. (1989). Thioredoxin and glutaredoxin systems. J Biol Chem 264, 13963-13966. Huang, S., Pettaway, C.A., Uehara, H., Bucana, C.D., and Fidler, I.J. (2001). Blockade of NF-kappaB activity in human prostate cancer cells is associated with suppression of angiogenesis, invasion, and metastasis. Oncogene 20, 4188-4197. Kaimul, A.M., Nakamura, H., Masutani, H., and Yodoi, J. (2007). Thioredoxin and thioredoxin-binding protein-2 in cancer and metabolic syndrome. Free Radic Biol Med 43, 861-868. Kokkola, A., Monni, O., Puolakkainen, P., Larramendy, M.L., Victorzon, M., Nordling, S., Haapiainen, R., Kivilaakso, E., and Knuutila, S. (1997). 17q12-21 amplicon, a novel recurrent genetic change in intestinal type of gastric carcinoma: a comparative genomic hybridization study. Genes Chromosomes Cancer 20, 38-43. Kute, T., Lack, C.M., Willingham, M., Bishwokama, B., Williams, H., Barrett, K., Mitchell, T., and Vaughn, J.P. (2004). Development of Herceptin resistance in breast cancer cells. Cytometry A 57, 86-93. Lu, J., and Holmgren, A. (2009). Selenoproteins. J Biol Chem 284, 723-727. Mahlamaki, E.H., Barlund, M., Tanner, M., Gorunova, L., Hoglund, M., Karhu, R., and Kallioniemi, A. (2002). Frequent amplification of 8q24, 11q, 17q, and 20q-specific genes in pancreatic cancer. Genes Chromosomes Cancer 35, 353-358. Maqani, N., Belkhiri, A., Moskaluk, C., Knuutila, S., Dar, A.A., and El-Rifai, W. (2006). Molecular dissection of 17q12 amplicon in upper gastrointestinal adenocarcinomas. Mol Cancer Res 4, 449-455. Nakamura, H., Nakamura, K., and Yodoi, J. (1997). Redox regulation of cellular activation. Annu Rev Immunol 15, 351-369. Ramanathan, R.K., Kirkpatrick, D.L., Belani, C.P., Friedland, D., Green, S.B., Chow, H.H., Cordova, C.A., Stratton, S.P., Sharlow, E.R., Baker, A., et al. (2007). A Phase I pharmacokinetic and pharmacodynamic study of PX-12, a novel inhibitor of thioredoxin-1, in patients with advanced solid tumors. Clin Cancer Res 13, 2109-2114. Ross, J.S., and Fletcher, J.A. (1999). The HER-2/neu oncogene: prognostic factor, predictive factor and target for therapy. Semin Cancer Biol 9, 125-138. Saitoh, M., Nishitoh, H., Fujii, M., Takeda, K., Tobiume, K., Sawada, Y., Kawabata, M., Miyazono, K., and Ichijo, H. (1998). Mammalian thioredoxin is a direct inhibitor of apoptosis signal-regulating kinase (ASK) 1. EMBO J 17, 2596-2606. Sinensky, M. (2000). Recent advances in the study of prenylated proteins. Biochim Biophys Acta 1484, 93-106. Tomasetto, C., Regnier, C., Moog-Lutz, C., Mattei, M.G., Chenard, M.P., Lidereau, R., Basset, P., and Rio, M.C. (1995). Identification of four novel human genes amplified and overexpressed in breast carcinoma and localized to the q11-q21.3 region of chromosome 17. Genomics 28, 367-376. Varis, A., Wolf, M., Monni, O., Vakkari, M.L., Kokkola, A., Moskaluk, C., Frierson, H., Jr., Powell, S.M., Knuutila, S., Kallioniemi, A., et al. (2002). Targets of gene amplification and overexpression at 17q in gastric cancer. Cancer Res 62, 2625-2629. Vinatzer, U., Dampier, B., Streubel, B., Pacher, M., Seewald, M.J., Stratowa, C., Kaserer, K., and Schreiber, M. (2005). Expression of HER2 and the coamplified genes GRB7 and MLN64 in human breast cancer: quantitative real-time reverse transcription-PCR as a diagnostic alternative to immunohistochemistry and fluorescence in situ hybridization. Clin Cancer Res 11, 8348-8357. Winter-Vann, A.M., and Casey, P.J. (2005). Post-prenylation-processing enzymes as new targets in oncogenesis. Nat Rev Cancer 5, 405-412. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23196 | - |
| dc.description.abstract | 癌症相關分子的發現，對於疾病的診斷與治療具有很大的應用潛力。C35 (C17orf37)是一個位於人類第十七號染色體(17q12-21)的新癌症相關基因。由於在前人利用免疫組織染色法（immunohistochemical analysis, IHC）所做的研究中指出，C35蛋白可大量且頻繁地在乳癌病理組織中被偵測到，因此在本實驗中，我們將從結構的分析與對細胞功能的影響，來探討C35蛋白在癌症生成中所扮演的角色。利用反轉錄作用從人類乳癌細胞—SK-BR-3所獲得的C35全長基因，在經過大腸桿菌（E. coli strain BL21）的表現後，我們將所純化的蛋白利用圓二色光譜儀（circular dichroism）和核磁共振儀（nuclear magnetic resonance）分析其結構。從圓二色光譜儀的分析結果顯示，C35的二級結構主要以α-螺旋（α-helix）的形式存在；此外，C35蛋白具有很好的熱耐受性和酸鹼穩定性。在利用核磁共振儀所做的測量中，首先我們利用一維分析法（one-dimensional NMR analysis）先確認C35具有穩定的蛋白構型，接著利用二維法（two-dimensional NMR analysis）對其三級結構做進一步地分析，最後，我們發現C35蛋白具有和硫氧還蛋白相似的構型（thioredoxin-like structure）。為了進一步瞭解C35蛋白和癌症的相關性，在接下來的研究中，我們利用免疫組織染色法，證實了C35也會在胃癌中表現；在反轉錄聚合酵素連鎖反應（RT-PCR）的偵測中，發現C35 mRNA 的表現普遍存在於所有的細胞中；然而在蛋白質的表現上，利用西方點墨法（Western blotting）偵測，我們只在人類乳癌細胞—BT474中，發現內生性C35蛋白的存在。為了探討C35蛋白在正常組織中所扮演的角色，我們利用10種不同的老鼠組織來做內生性C35蛋白的偵測。由於在這些正常組織中，都無法偵測到C35蛋白的存在，因此我們推測C35蛋白並不會在正常組織中表現。最後，我們利用Boyden chamber assay探討C35蛋白對細胞（CHO-K1）功能的影響，實驗結果顯示，C35的大量表現具有促進細胞移動的能力。藉由C35蛋白的初步研究，顯示其與癌症的發生確實有相關性，然而其在癌化過程中所扮演的角色，仍有待更進一步的研究。 | zh_TW |
| dc.description.abstract | Identification of cancer-specific biomarkers has enormous potential to enhance detection, treatment, and prognosis. C35 (C17orf37) is a novel gene, which locates at chromosome 17q12-21 and is found to be up-regulated in many cancers. In a previous study, immunohistochemical analysis detected robust and frequent expression of C35 protein in breast cancer tissues. Here, we attempt to characterize the properties of C35 protein as well as to demonstrate a functional association between C35 and tumor progression. We cloned the full length gene of C35 from a breast cancer cell line, SK-BR-3, and expressed it to gain recombinant C35 protein from E. coli for circular dichroism(CD) and nuclear magnetic resonance (NMR) analyses. The CD spectrum of C35 protein revealed a possible alpha-helix secondary conformation existed. To our surprise, we also observed an extremely heat tolerance and a pH stable properties of this protein. Additionally, based on one-dimensional NMR analysis, it also showed a well organize protein structure. Finally, tertiary structure determined by two-dimensional NMR assignment reveals C35 as thioredoxin-like molecule. To warrant C35 as a cancer related gene, we first survey its expression in different cancer cell lines and tissues by using RT-PCR, Western blot and immunohistochemistry, respectively. Immunohistochemistry had showed high relevance of C35 in gastric cancer. Otherwise, survey of different cell lines by Western blotting reveals highly endogenous C35 expression in breast cancer cells, BT474. Furthermore, to investigate the function of C35 in normal tissues, analyzing of ten normal mice’s tissues reveals no detectable C35 protein. Effects of C35 on cellular proliferation, migration and apoptosis are investigating. In a preliminary test using the modified Boyden chamber assay, overexpression of C35 enables promoting cell migration toward fetal bovine serum. Beyond this, we will further establish the role of C35 in tumor progression. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T04:46:54Z (GMT). No. of bitstreams: 1 ntu-98-R96633003-1.pdf: 2162405 bytes, checksum: 5b75ab0a0cf338e559c134e94fe3172f (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | Abstract in Chinese…………………………………………..…………….….…..… I
Abstract in English…………………………………………..…………….….…….III Contents……………...……………………………………………………………….V List of Figures……...…………………………………………………………..…...VII INTRODUCTION……………………………………………………………………1 1. Cancer……………………………………………………………………….1 2. Human chromosome 17q12-q21……………………………………………1 3. C35 (C17orf37) is a novel tumor biomarker………………………………..2 MATERIALS and METHODS………………………….………………………......5 1. Antibodies…….……………………………………………..……….….......5 2. RNA extraction and reverser transcription-PCR………...…...……………..5 3. Construction of expression plasmids………………………………………..6 4. Preparation of purified C35 protein…………...………………………….....7 5. Cell lines and culture condition…………………….……………….............8 6. Transfection procedure…………………….………………..……………....8 7. Immunoprecipitation and Western blotting analyses...……………………..9 8. Immunohistochemistry………………………………………………….....10 9. Cell migration assay…………………………………………………...…..11 RESULTS……………………………………………………………………………12 1. Construction of a novel gene, C35 (C17orf37)……………………...…….12 2. Structure determination of C35……………………………………………12 2.1 Circular dichroism (CD)……………………………………………...12 2.2 Nuclear magnetic resonance (NMR)…………………………………13 2.3 Crystallization………………………………………………………..14 3. Protein properties of C35………………………………………………….15 4. Generation of anti-C35 antibody…………………………………………..15 5. C35 protein detection by immunohistochemical staining……...………….16 6. C35 expression analyzed by reverse transcription (RT)–PCR…………….17 7. Endogenous C35 protein surveyed by Western blotting analysis……...….18 8. Scanning of mouse tissues for C35………………………………………..18 9. Cell migration assay……………………………………………….………19 DISCUSSION ……………………………………………...……………………….20 FIGURES……………………………………………………………………….…...24 REFERENCES…………………………………………………………………..….55 APPENDIX………………………………………………………………………….60 | |
| dc.language.iso | en | |
| dc.subject | thioredoxin | zh_TW |
| dc.subject | cancer biomarker | zh_TW |
| dc.subject | C35 (C17orf37) | zh_TW |
| dc.subject | chromosome 17q12-21 | zh_TW |
| dc.subject | nuclear magnetic resonance | zh_TW |
| dc.title | 癌症相關蛋白C35 (C17orf37)的結構、生化以及臨床分析 | zh_TW |
| dc.title | Structural, biochemical and clinical analyses of a cancer-related protein, C35 (C17orf37) | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 徐駿森,黃敏銓,賴逸儒 | |
| dc.subject.keyword | cancer biomarker,C35 (C17orf37),chromosome 17q12-21,nuclear magnetic resonance,thioredoxin, | zh_TW |
| dc.relation.page | 62 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2009-07-30 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 植物病理與微生物學研究所 | zh_TW |
| 顯示於系所單位： | 植物病理與微生物學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-98-1.pdf 未授權公開取用 | 2.11 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。