請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/19446
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 吳漢忠(Han-Chung Wu) | |
dc.contributor.author | Chien-Hsu Chen | en |
dc.contributor.author | 陳建旭 | zh_TW |
dc.date.accessioned | 2021-06-08T01:59:26Z | - |
dc.date.copyright | 2016-08-26 | |
dc.date.issued | 2016 | |
dc.date.submitted | 2016-06-24 | |
dc.identifier.citation | 1. Nardi, N. Beyer; da Silva Meirelles, L. (2006). 'Mesenchymal Stem Cells: Isolation, In Vitro Expansion and Characterization'. In Wobus, Anna M.; Boheler, Kenneth. Stem Cells 174, 249–82
2. Visvader, J.E., and Lindeman, G.J. (2008). Cancer stem cells in solid tumors: accumulating evidence and unresolved questions. Nature reviews Cancer 8, 755-768 3. Litvinov, S. V., Velders, M. P., Bakker, H. A., Fleuren, G. J., and Warnaar, S. O. (1994) Ep-CAM: a human epithelial antigen is a homophilic cell-cell adhesion molecule. The Journal of cell biology 125, 437-446. 4. Momburg, F., Moldenhauer, G., Hammerling, G. J., and Moller, P. (1987) Immunohistochemical study of the expression of a Mr 34,000 human epithelium-specific surface glycoprotein in normal and malignant tissues. Cancer research 47, 2883-2891. 5. Maetzel, D., Denzel, S., Mack, B., Canis, M., Went, P., Benk, M., Kieu, C., Papior, P., Baeuerle, P. A., Munz, M., and Gires, O. (2009) Nuclear signalling by tumour-associated antigen EpCAM. Nature cell biology 11, 162-171 6. Maghzal, N., Vogt, E., Reintsch, W., Fraser, J. S., and Fagotto, F. (2010) The tumor-associated EpCAM regulates morphogenetic movements through intracellular signaling. Journal of Cell Biology 191, 645-659 7. Stoyanova, T., Goldstein, A. S., Cai, H., Drake, J. M., Huang, J., and Witte, O. N. (2012) Regulated proteolysis of Trop2 drives epithelial hyperplasia and stem cell self-renewal via beta-catenin signaling. Genes & development 26, 2271-2285 8. Munz, M., Kieu, C., Mack, B., Schmitt, B., Zeidler, R., and Gires, O. (2004) The carcinoma-associated antigen EpCAM upregulates c-myc and induces cell proliferation. Oncogene 23, 5748-5758 9. Pittenger (1999). Multilineage potential of adult human mesenchymal stem cells. Science 284 (5411): 143–147 10. Brighton CT, Hunt RM (1997). Early histologic and ultrastructural changes in micro-vessels of periosteal callus. J Orthop Trauma 11 (4): 244–53 11. Valero MC, Huntsman HD, Liu J, Zou K, Boppart MD (2012). Eccentric exercise facilitates mesenchymal stem cell appearance in skeletal muscle. PLoS ONE 7 (1): 29760 12. Wang S, et al. (2012). Clinical applications of mesenchymal stem cells. JOURNAL OF HEMATOLOGY & ONCOLOGY 5 (19) 13. Briggs R, King TJ. Transplantation of living nuclei from blastula cells into enucleated frogs eggs. Proc Natl Acad Sci USA (38): 455 14. Gurdon JB. The developmental capacity of nuclei taken from intestinal epithelium cells of feeding tadpoles. Journal of embryology and experimental morphology (10): 622–640 15. Muenthaisong, S., Ujhelly, O., Polgar, Z., Varga, E., Ivics, Z., Pirity, M. K., and Dinnyes, A. (2012) Generation of mouse induced pluripotent stem cells from different genetic backgrounds using Sleeping beauty transposon 16. Davis, R. P., Nemes, C., Varga, E., Freund, C., Kosmidis, G., Gkatzis, K., de Jong, D., Szuhai, K., Dinnyes, A., and Mummery, C. L. (2013) Generation of induced pluripotent stem cells from human foetal fibroblasts using the Sleeping Beauty transposon gene delivery system. Differentiation; research in biological diversity 86, 30-37 17. Klincumhom, N., Pirity, M. K., Berzsenyi, S., Ujhelly, O., Muenthaisong, S., Rungarunlert, S., Tharasanit, T., Techakumphu, M., and Dinnyes, A. (2012) Generation of neuronal progenitor cells and neurons from mouse sleeping beauty transposon-generated induced pluripotent stem cells. Cellular reprogramming 14, 390-397 18. Okita, K., Nakagawa, M., Hyenjong, H., Ichisaka, T., and Yamanaka, S. (2008) Generation of mouse induced pluripotent stem cells without viral vectors. Science 322, 949-953 19. Stadtfeld, M., Nagaya, M., Utikal, J., Weir, G., and Hochedlinger, K. (2008) Induced pluripotent stem cells generated without viral integration. Science 322, 945-949 20. Zhou, W., and Freed, C. R. (2009) Adenoviral gene delivery can reprogram human fibroblasts to induced pluripotent stem cells. Stem cells 27, 2667-2674 21. Kim, D., Kim, C. H., Moon, J. I., Chung, Y. G., Chang, M. Y., Han, B. S., Ko, S., Yang, E., Cha, K. Y., Lanza, R., and Kim, K. S. (2009) Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins. Cell stem cell 4, 472-476 22. Zhang, H., Ma, Y., Gu, J., Liao, B., Li, J., Wong, J., and Jin, Y. (2012) Reprogramming of somatic cells via TAT-mediated protein transduction of recombinant factors. Biomaterials 33, 5047-5055 23. Fusaki, N., Ban, H., Nishiyama, A., Saeki, K., and Hasegawa, M. (2009) Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome. Proceedings of the Japan Academy. Series B, Physical and biological sciences 85, 348-362 24. Ban, H., Nishishita, N., Fusaki, N., Tabata, T., Saeki, K., Shikamura, M., Takada, N., Inoue, M., Hasegawa, M., Kawamata, S., and Nishikawa, S. (2011) Efficient generation of transgene-free human induced pluripotent stem cells (iPSCs) by temperature-sensitive Sendai virus vectors. Proceedings of the National Academy of Sciences of the United States of America 108, 14234-14239 25. Warren, L., Manos, P. D., Ahfeldt, T., Loh, Y. H., Li, H., Lau, F., Ebina, W., Mandal, P. K., Smith, Z. D., Meissner, A., Daley, G. Q., Brack, A. S., Collins, J. J., Cowan, C., Schlaeger, T. M., and Rossi, D. J. (2010) Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell stem cell 7, 618-630 26. Hiratsuka, M., Uno, N., Ueda, K., Kurosaki, H., Imaoka, N., Kazuki, K., Ueno, E., Akakura, Y., Katoh, M., Osaki, M., Kazuki, Y., Nakagawa, M., Yamanaka, S., and Oshimura, M. (2011) Integration-free iPS cells engineered using human artificial chromosome vectors. Plos One 6, e25961 27. Gordon, M. Y. (2008) Stem cells for regenerative medicine--biological attributes and clinical application. Experimental hematology 36, 726-732 28. Corsten, M. F., and Shah, K. (2008) Therapeutic stem-cells for cancer treatment: hopes and hurdles in tactical warfare. The lancet oncology 9, 376-384 29. Keller, G. (2005) Embryonic stem cell differentiation: emergence of a new era in biology and medicine. Genes & development 19, 1129-1155 30. Takahashi, K., and Yamanaka, S. (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663-676 31. Park, I. H., Zhao, R., West, J. A., Yabuuchi, A., Huo, H., Ince, T. A., Lerou, P. H., Lensch, M. W., and Daley, G. Q. (2008) Reprogramming of human somatic cells to pluripotency with defined factors. Nature 451, 141-146 32. Short, B. J., Brouard, N., and Simmons, P. J. (2009) Prospective isolation of mesenchymal stem cells from mouse compact bone. Methods Mol Biol 482, 259-268 33. Pittenger, M. F., Mackay, A. M., Beck, S. C., Jaiswal, R. K., Douglas, R., Mosca, J. D., Moorman, M. A., Simonetti, D. W., Craig, S., and Marshak, D. R. (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284, 143-147 34. Prall, W. C., Haasters, F., Heggebo, J., Polzer, H., Schwarz, C., Gassner, C., Grote, S., Anz, D., Jager, M., Mutschler, W., and Schieker, M. (2013) Mesenchymal stem cells from osteoporotic patients feature impaired signal transduction but sustained osteoinduction in response to BMP-2 stimulation. Biochemical and biophysical research communications 440, 617-622 35. Wu, J., Wu, K., Lin, F., Luo, Q., Yang, L., Shi, Y., Song, G., and Sung, K. L. (2013) Mechano-growth factor induces migration of rat mesenchymal stem cells by altering its mechanical properties and activating ERK pathway. Biochemical and biophysical research communications 441, 202-207 36. Dombrowski, C., Helledie, T., Ling, L., Grunert, M., Canning, C. A., Jones, C. M., Hui, J. H., Nurcombe, V., van Wijnen, A. J., and Cool, S. M. (2013) 37. Huang, H. P., Chen, P. H., Yu, C. Y., Chuang, C. Y., Stone, L., Hsiao, W. C., Li, C. L., Tsai, S. C., Chen, K. Y., Chen, H. F., Ho, H. N., and Kuo, H. C. (2011) Epithelial cell adhesion molecule (EpCAM) complex proteins promote transcription factor-mediated pluripotency reprogramming. J Biol Chem 286, 33520-33532 38. Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KHS. Viable offspring derived from fetal and adult mammalian cells. Nature 1997; 385: 810-813 39. Sei-Myoung Han1, Sang-Hun Han1, Ye-Rin Coh1, Goo Jang2, Jeong Chan Ra3, Sung-Keun Kang3, Hee-Woo Lee4 and Hwa-Young Youn. Enhanced proliferation and differentiation of Oct4- and Sox2-overexpressing human adipose tissue mesenchymal stem cells. Experimental & Molecular Medicine 2014; 46 40. Tada M, Tada T, Lefebvre L, Barton SC and Surani MA. Embryonic germ cells induce epigenetic reprogramming of somatic nucleus in hybrid cells. EMBO J. 1997; 16: 6510-6520 41. Takahashi K and Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006; 126 (4): 663–76 42. Speman H. Embryonic development and induction. New Haven: Yale University press 1938 43. Lin, C.W., Liao, M.Y., Lin, W.W., Wang, Y.P., and Wu, H.C. (2012). Epithelial cell adhesion molecule regulates tumor initiation and tumorigenesis via activating reprogramming factors and epithelial-mesenchymal transition gene expression in colon cancer. The Journal of biological chemistry 287, 39449-39459 44. Yu, K. R., Yang, S. R., Jung, J. W., Kim, H., Ko, K., Han, D. W., Park, S. B., Choi, S. W., Kang, S. K., Scholer, H., and Kang, K. S. (2012) CD49f enhances multipotency and maintains stemness through the direct regulation of OCT4 and SOX2. Stem cells 30, 876-887 45. Satomura, K., Derubeis, A. R., Fedarko, N. S., Ibaraki-O'Connor, K., Kuznetsov, S. A., Rowe, D. W., Young, M. F., and Gehron Robey, P. (1998) Receptor tyrosine kinase expression in human bone marrow stromal cells. Journal of cellular physiology 177, 426-438 46. Normanno, N., De Luca, A., Aldinucci, D., Maiello, M. R., Mancino, M., D'Antonio, A., De Filippi, R., and Pinto, A. (2005) Gefitinib inhibits the ability of human bone marrow stromal cells to induce osteoclast differentiation: implications for the pathogenesis and treatment of bone metastasis. Endocrine-related cancer 12, 471-482 47. Zhu, J., Jia, X., Xiao, G., Kang, Y., Partridge, N. C., and Qin, L. (2007) EGF-like ligands stimulate osteoclastogenesis by regulating expression of osteoclast regulatory factors by osteoblasts: implications for osteolytic bone metastases. J Biol Chem 282, 26656-26664 48. Han, W., and Lo, H. W. (2012) Landscape of EGFR signaling network in human cancers: biology and therapeutic response in relation to receptor subcellular locations. Cancer letters 318, 124-134 49. Satomura, K., Derubeis, A. R., Fedarko, N. S., Ibaraki-O'Connor, K., Kuznetsov, S. A., Rowe, D. W., Young, M. F., and Gehron Robey, P. (1998) Receptor tyrosine kinase expression in human bone marrow stromal cells. Journal of cellular physiology (177): 426-438 50. Yamanaka, S. (2012) Induced pluripotent stem cells: past, present, and future. Cell stem cell (10): 678-684 51. Tung-Ying Lu, Ruei-Min Lu, Mei-Ying Liao, John Yu, Chu-Hung Chung, Cheng-Fu Kao, and Han-Chung Wu. (2010) Epithelial Cell Adhesion Molecule Regulation Is Associated with the Maintenance of the Undifferentiated Phenotype of Human Embryonic Stem Cells. The Journal of Biological Chemistry 285, 8719-8732 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/19446 | - |
dc.description.abstract | 人類間質幹細胞可以分化成多種細胞類型,包括:軟骨細胞、成骨細胞、肌細胞及脂肪細胞。上皮細胞黏附因子(EpCAM),屬於第一型穿膜醣蛋白,在所有的腺狀細胞癌以及鱗狀上皮細胞癌中都有很高的表現量。並且是種很常見的癌症幹細胞標定物,也會促使惡性腫瘤的生長。近年來也發現EpCAM 於未分化的細胞中有高度表現,且在已經分化的胚胎幹細胞中表達較低,EpCAM 的蛋白和轉錄水平在,人類胚胎幹細胞一開始分化就會迅速減弱表現,後來研究發現EpCAM 可作為幹細胞的重要表面標記物。我們先前的研究發現EpCAM、EpICD 能直接接上幾個基因promoter,其中包括c-Myc,Oct4,Nanog,Sox2,Klf4,以幫助維持胚胎幹細胞的多能性。因此,我們推測EpCAM 可能在重編程的iPSCs 多能性,且有重要功能。在我們的研究中,也發現EpCAM 會表現在臍帶血間質幹細胞表並且EpEX 能促進臍帶血間質幹細胞和骨髓間質幹細胞的增生基因的表現。此外,我們還發現EpEX 可以促進細胞增生是透過增加STAT3 磷酸化和提高細胞週期調節因子的表現,並且是透過EGF 的接受器來影響到增生基因Lin28 的表現和STAT3 的磷酸化。因此,EpEX 不僅提高幹細胞自我修復、增生能力,也能夠在幹細胞重編程和多能性維持也起到重要作用。根據我們的研究,我們將提供EpCAM 的深入調控機制,有助於開發更高效的重編程方法以及策略,讓幹細胞維持iPS 的細胞形態以及細胞多能性的維護。 | zh_TW |
dc.description.abstract | Mesenchymal stem cells, or MSCs, are multipotent stromal cells that can differentiate into a variety of cell types, including chondrocytes (cartilage cells), osteoblasts (bone cells), myocytes (muscle cells), and adipocytes (fat cells) (1). The epithelial cell adhesion molecule (EpCAM) is a type I transmembrane glycoprotein overexpressed in various carcinomas, such as adenocarcinomas and squamous cell carcinomas. Recently, EpCAM has been found as a well-known cancer stem cell marker and contributes to tumor growth. We have shown that EpCAM is highly expressed by undifferentiated rather than differentiated hESCs. In addition, EpCAM directly binds to the promoter of several reprogramming genes, including c-Myc, Oct4, Nanog, Sox2, and Klf4, to help maintain the pluripotency of hESCs, suggesting that in addition to being a potent stem cell marker, EpCAM also plays a crucial role in hESCs reprogramming and pluripotency maintenance. In the present study, we found that EpCAM is expressed in cord blood MSCs, and EpEX is also found to up-regulate the proliferation gene expressions in both cord blood MSCs and bone marrow MSCs. We also found that EpEX increases STAT3 phosphorylation, Lin28 protein level through EGF receptor and that phospho-STAT3 can interact with Lin28 to promote cell proliferation and cell cycle progression. Additionally, EpEX induces the expression of cell cycle regulators. We found that EpEX can serve as a cytokine and plays crucial roles in MSCs reprogramming a pluritpotency maintenance. Based on our study, we provided the molecular mechanism of EpCAM, in the regulation of stemness of MSCs. This results suggest that the stemness of MSCs and iPSCs can be further enhanced through EpCAM signaling. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T01:59:26Z (GMT). No. of bitstreams: 1 ntu-105-R03450005-1.pdf: 4600088 bytes, checksum: 77fe493bad11496fdff031488f1700c1 (MD5) Previous issue date: 2016 | en |
dc.description.tableofcontents | 誌謝………………………………………..…………………………………………….1
中文摘要………………………………………………………………………………...3 Abstract…………………………………………………………………………………4 Table of Contents……………………………………………………………………….6 List of abbreviations..…………………………………………………………………..9 Introduction……………………………………………………………………………11 1.1 Epithelial cell adhesion molecule (EpCAM)…………………………………..11 1.2 Mesenchymal stem cell………………………………………………………13 1.3 Induced pluripotent stem cells…………………………………………………14 1.4 Expression of EpCAM in stem cells, progenitor cells and MSCs……………16 1.5 The epidermal growth factor receptor (EGFR) pathways………………………17 1.6 Specific aims…………………………………………………………………19 Materials and Methods………………...……………………………………………22 2.1 Cell lines and culture condition………………………………………..………22 2.2 Western blot analysis……………………………………………………….….22 2.3 Flow Cytometry Analysis……………………………………………………...23 2.4 Plasmids and Lentivirus Preparation……………………………………..……23 2.5 RNA extraction and purification…………………………………………...….25 2.6 Reverse transcription PCR……………………………………………….……25 2.7 Quantitative reverse transcription polymerase chain reaction (qRT-PCR)…….26 2.8 Sphere formation assays……………………………………………………….27 2.9 In situ hybridization……………………………………………………………27 2.10 Statistical analysis…………………………………………………………....30 2.11 Immunoprecipitation and immunoblotting…………………………………30 Results…………………...……………………………………………………………32 3.1 EpEX enhances the cell proliferation and self-renewal in mesenchymal stem cells…………………………………………………………………………….….32 3.2 EpEX induces the phosphorylation of EGFR…………………………34 3.3 EpEX enhances cell proliferation and multipotency through EGFR signal………………………………………………………….……………………35 3.4 EpEX induces cell proliferation and self-renewal via STAT3 signal…36 3.5 EpEX induces STAT3 signal through EGFR…………………………...……38 3.6 EpEX attenuates miRNA Let-7……………………………………..……….38 3.7 EpEX attenuates microRNA Let-7 through EGFR-STAT3……………….…39 3.8 Knocking down expression up-regulates microRNA Let-7 expression in CB MSCs and BM MSCs………………………………………………………….…..40 3.9 Downregulation of EpCAM decreases the proliferation and stemness markers of mesenchymal stem cells through STAT3 signaling……………………………..40 3.10 EpEX increases phophoSTAT3 to interact with Lin28………………………41 3.11 Time course of EpEX enhances cell proliferation and pluripotency through EGFR signal……………………………………………………………………….43 3.12 Time course of EpEX enhances cell proliferation and pluripotency through STAT3 signal………………………………………………………………………43 3.13 Knocking down EGFR decreases the proliferation and stemness of mesenchymal stem cells through STAT3 signaling………………………………..44 3.14 Knocking down STAT3 decreases the proliferation and stemness of mesenchymal stem cells…………………………………………………...………45 3.15 EGFR and STAT3 abolished up-regulates microRNA Let-7 expression in CB MSCs and BM MSCs…………………………………………………..………….45 Discussion………………………………………………………………...…………...47 References……………………………………………………………………………...96 Appendix……………………………………………………………………………105 | |
dc.language.iso | en | |
dc.title | 上皮細胞黏附因子在間質幹細胞調控活化與訊息傳遞
的分子機制 | zh_TW |
dc.title | The Molecular Mechanisms of Epithelial Cell Adhesion
Molecule (EpCAM) in Regulation of Mesenchymal Stem Cells Signaling | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 呂仁(Jean Lu),沈家寧(Chia-Ning Shen),郭紘志(Hung-Chih Kuo) | |
dc.subject.keyword | 間質幹細胞,上皮細胞黏附因子,上皮細胞黏附因子之細胞外域,上皮細胞黏附因子之細胞內域,EGFR,STAT3,Lin28,Let-7e, | zh_TW |
dc.subject.keyword | Mesenchymal stem cell,Epithelial cell adhesion molecule,Extracellular domain of EpCAM,Intracellular domain of EpCAM,EGFR,STAT3,Lin28,Let-7e, | en |
dc.relation.page | 105 | |
dc.identifier.doi | 10.6342/NTU201600450 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2016-06-24 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 口腔生物科學研究所 | zh_TW |
顯示於系所單位: | 口腔生物科學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-105-1.pdf 目前未授權公開取用 | 4.49 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。