請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62952
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 朱有田(Yu-Ten Ju) | |
dc.contributor.author | Ching-Ying Huang | en |
dc.contributor.author | 黃瀞瑩 | zh_TW |
dc.date.accessioned | 2021-06-16T16:16:13Z | - |
dc.date.available | 2017-12-31 | |
dc.date.copyright | 2013-03-06 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-02-05 | |
dc.identifier.citation | Chapter 1
Band, V. 1998. The role of retinoblastoma and p53 tumor suppressor pathways in human mammary epithelial cell immortalization. J. Oncol. 12:499-507. Boutinaud, M., and H. Jammes. 2002. Potential uses of milk epithelial cells: a review. Reprod. Nutr. Dev. 42:133-147. Boyer, S. N., D. E. Wazer, and V. Band. 1996. E7 protein of human papilloma virus-16 induces degradation of retinoblastoma protein through the ubiquitin-proteasome pathway. Cancer Res. 56:4620-4624. Clark, A. J. 1998. The mammary gland as a bioreactor: expression, processing, and production of recombinant proteins. J. Mammary Gland Biol. Neoplasia 3:337–350. Clark, A. J., J. P. Simons, and I. Wilmut. 1992. Germline manipulation: applications in agriculture and biotechnology. Pages 247-269 in Transgenic Mice in Biology and Medicine. F. Grosveld and G. Kollias, ed. Academic Press, London, UK. Clarkson, R. W., M. T. Wayland, J. Lee, T. Freeman, and C. J. Watson. 2004. Gene expression profiling of mammary gland development reveals putative roles for death receptors and immune mediators in postlactational regression. Breast Cancer Res. 6:R92-109. Debnath, J., K. R. Mills, N. L. Collins, M. J. Reginato, S. K. Muthuswamy, and J. S. Brugge. 2002. The role of apoptosis in creating and maintaining luminal space within normal and oncogene-expressing mammary acini. Cell 111:29-40. Debnath, J., S. K. Muthuswamy, and J. S. Brugge. 2003. Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures. Methods 30:256-268. Devinoy, E., D. Thepot, M. G. Stinnakre, M. L. Fontaine, H. Grabowski, C. Puissant, A. Pavirani, and L. M. Houdebine. 1994. High level production of human growth hormone in the milk of transgenic mice: the upstream region of the rabbit whey acidic protein (WAP) gene targets transgene expression to the mammary gland. Transgenic Res. 3:79-89. Edmunds, T., S. M. Van Patten, J. Pollock, E. Hanson, R. Bernasconi, E. Higgins, P. Manavalan, C. Ziomek, H. Meade, J. M. McPherson, and E. S. Cole. 1998. Transgenically produced human antithrombin: structural and functional comparison to human plasma-derived antithrombin. Blood 91:4561-4567. Fan, W., K. Plaut, A. J. Bramley, J. W. Barlow, S. A. Mischler, and D. E. Kerr. 2004. Persistency of adenoviral-mediated lysostaphin expression in goat mammary glands. J. Dairy Sci. 87: 602-608. Gewin, L., H. Myers, T. Kiyono, and D. A. Galloway. 2004. Identification of a novel telomerase repressor that interacts with the human papillomavirus type-16 E6/E6-AP complex. Genes Dev. 18:2269-2282. Hennighausen, L., and G. W. Robinson. 2001. Signaling pathways in mammary gland development. Dev. Cell 1:467-475. Hinck. L., and G. B. Silberstein. 2005. Key stages in mammary gland development: the mammary end bud as a motile organ. Breast Cancer Res. 7:245-251. Hovey, R. C., J. F. Trott, and B. K. Vonderhaar. 2002. Establishing a framework for the functional mammary gland: from endocrinology to morphology. J. Mammary Gland Biol. Neoplasia 7:17-38. Itahana, K., J. Campisi, and G. P. Dimri. 2004. Mechanisms of cellular senescence in human and mouse cells. Biogerontology 5:1-10. Ke, M. W., J. T. Hsu, Y. N. Jiang, W. T. K. Cheng, Y. T. Ju. 2011. Characterization of hTERT-immortalized caprine mammary epithelial cells. Reprod. Dom. Anim. doi: 10.1111/j.1439-0531.2011.01916.x. Kiyono, T., S. A. Foster, J. I. Koop, J. K. McDougall, D. A. Galloway, and A. J. Klingelhutz. 1998. Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cells. Nature 396: 84-88. Medina, D. 1996. The mammary gland: A unique organ for the study of development and tumorigenesis. J. Mammary Gland Biol. Neoplasia 1:5-19. Moura, R. R., L. M. Melo, and V. J. F. Freitas. 2011. Production of recombinant proteins in milk of transgenic and non-transgenic goats. Braz. Arch. Biol. Technol. 54:927-938. Muthuswamy, S. K., D. Li, S. Lelievre, M. J. Bissell, and J. S. Brugge. 2001. ErbB2, but not ErbB1, reinitiates proliferation and induces luminal repopulation in epithelial acini. Nat. Cell Biol. 3:785-792. Pantschenko, A. G., J. Woodcocl-Mitchell, S. L. Bushmich, and T. J. Yang. 2000 Establishment and characterization of a caprine mammary epithelial cell line (CMEC). In Vitro Cell Dev. Biol. Anim. 36:351-356. Ratsch, S. B., Q. Gao, S. Srinivasan, D. E. Wazer, and V. Band. 2001. Multiple genetic changes are required for efficient immortalization of different subtypes of normal human mammary epithelial cells. Radiat. Res. 155:143-150. Shay, J. W., B. A. Van Der Haegen, Y. Ying, and W. E. Wright. 1993. The frequency of immortalization of human fibroblasts and mammary epithelial cells transfected with SV40 large T-antigen. Exp. Cell Res. 209:45-52. Shay, J. W., and W. E. Wright. 2005. Senescence and immortalization: role of telomeres and telomerase. Carcinogenesis 26:867-874. Sternlicht, M. D. 2006. Key stages in mammary gland development: the cues that regulate ductal branching morphogenesis. Breast Cancer Res. 8:201 (doi:10.1186/bcr1368). Sutherland, K. D., G. J. Lindeman, and J. E. Visvader. 2007. The molecular culprits underlying precocious mammary gland involution. J. Mammary Gland Biol. Neoplasia. 12:15-23. Toouli, C. D., L. I. Huschtscha, A. A. Neumann, J. R. Noble, L. M. Colgin, B. Hukku, and R. R. Reddel. 2002. Comparison of human mammary epithelial cells immortalized by simian virus 40 T-Antigen or by the telomerase catalytic subunit. Oncogene 21: 128-139. Varki, A. 1993. Biological roles of oligosaccharides: all of the theories are correct. Glycobiology 3:97-130. Wazer, D. E., X. L. Liu, Q. Chu, Q. Gao, and V. Band. 1995. Immortalization of distinct human mammary epithelial cell types by human papilloma virus 16 E6 or E7. Proc. Natl. Acad. Sci. U. S. A. 92:3687-3691. Wooding, F. B., G. Morgan, and H. Craig. 1977. “Sunbursts” and “christiesomes”: cellular fragments in normal cow and goat milk. Cell Tissue Res. 185:535-545. Woodworth, C. D., J. Chung, E. McMullin, G. D. Plowman, S. Simpson, and M. Iglesias. 1996. Transforming growth factor beta 1 supports autonomous growth of human papillomavirus-immortalized cervical keratinocytes under conditions promoting squamous differentiation. Cell Growth Differ. 7:811-820. Wu, H. T., C. S. Lin, and M. C. Huang. 2003. In vitro and ex vivo green fluorescent protein expression in alveolar mammary epithelial cells and mammary glands driven by the distal 5’-regulative sequence and intron 1 of the goat beta-casein gene. Reprod. Fertil. Dev. 15:231-239. Chapter 2 Baba, T., and O. Schneewind. 1996. Target cell specificity of a bacteriocin molecule: a C-terminal signal directs lysostaphin to the cell wall of Staphylococcus aureus. EMBO J. 15:4789-4797. Barkema, H. W., Y. H. Schukken, T. J. Lam, M. L. Beiboer, H. Wilmink, G. Benedictus, and A. J. Brand. 1998. Incidence of clinical mastitis in dairy herds grouped in three categories by bulk milk somatic cell counts. Dairy Sci. 81:411-149. Blom, N., T. Sicheritz-Ponten, R. Gupta, S. Gammeltoft, and S. Brunak. 2004. Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. Proteomics 4:1633-1649. Brouillette, E., G. Grondin, L. Shkreta, P. Lacasse, and B. G. Talbot. 2003. In vivo and in vitro demonstration that Staphylococcus aureus is an intracellular pathogen in the presence or absence of fibronectin-binding proteins. Microb. Pathog. 35:159-168. Burgoyne, R. D., and J. S. Duncan. 1998. Secretion of milk proteins. J. Mammary Gland Biol. Neoplasia 3:275-286 De Oliveira, A. P., J. L. Watts, S. L. Salmon, and F. M. Aarestrup. 2000. Antimicrobial susceptibility of Staphylococcus aureus isolated from bovine mastitis in Europe and the United States. J. Dairy Sci. 83:855–862. Dodd, F. H., and E. R. Jackson. (eds) 1971. Control of Bovine Mastitis. UK: British Cattle Veterinary Association. Dodd, F. H. and F. K. Neave. 1970. Progress in Mastitis Control. Shinfield, Reading: Biennial Reviews of the National Institute for Research in Dairying. p21. Doud, S. K., M. M. Chou, and D. A. Kendall. 1993. Titration of protein transport activity by incremental changes in signal peptide hydrophobicity. Biochemistry 32:1251-1256. Esslemont, D., and M. Kossaibati. 2002. Mastitis: how to get out of the dark ages. Vet. J. 164:85-86. Fan, W., K. Plaut, A. J. Bramley, J. W. Barlow, and D. E. Kerr. 2002. Adenoviral-mediated transfer of a lysostaphin gene into the goat mammary gland. J. Dairy Sci. 85:1709-1716. Fan, W., K. Plaut, A. J. Bramley, J. W. Barlow, S. A. Mischler, and D. E. Kerr. 2004. Persistency of adenoviral-mediated lysostaphin expression in goat mammary glands. J. Dairy Sci. 87: 602-608. Gargis, S. R., H. E. Heath, P. A. LeBlanc, L. Dekker, R. S. Simmonds, and G. L. Sloan. 2010. Inhibition of the activity of both domains of lysostaphin through peptidoglycan modification by the lysostaphin immunity protein. Appl. Environ. Microbiol. 76:6944-6946. Gruet, P., P. Maincent, X. Berthelot, and V. Kaltsatos. 2001. Bovine mastitis and intramammary drug delivery: review and perspectives. Adv. Drug Deliv. Rev. 50:245-259. Hall, J., G. P. Hazlewood, M. A. Surani, B. H. Hirst, and H. J. Gilbert. 1990. Eukaryotic and prokaryotic signal peptides: direct secretion of a bacterial endoglucanase by mammalian cells. J. Biol. Chem. 265:19996-19999. Hamby, S. E., and J. D. Hirst. 2008. Prediction of glycosylation sites using random forests. BMC Bioinformatics 9:500. Hatsuzawa, K., M. Tagaya, and S. Mizushima. 1997. The hydrophobic region of signal peptides is a determinant for SRP recognition and protein translocation across the ER membrane. J. Biochem.121:270-277. Hillerton, J. E., and E. A. Berry. 2005. Treating mastitis in the cow-a tradition or an archaism. J. Appl. Microbiol. 98:1250-1255. Holmberg, M., W. F. Fikse, L. Andersson-Eklund, K. Artursson, and A. Lunden. 2012. Genetic analyses of pathogen-specific mastitis. J. Anim. Breed. Genet. 129:129-137. Hounsell, E. F., M. J. Davies, and D. V. Renouf. 1996. O-linked protein glycosylation structure and function. Glycoconj. J. 13:19-26. Izard, J. W., M. B. Doughty, and D. A. Kendall. 1995. Physical and conformational properties of synthetic idealized signal sequences parallel their biological function. Biochemistry 34:9904-9912. Jenness, R. 1980. Composition and characteristics of goat milk: review 1968-1979. J. Dairy Sci. 63: 1605-1630. Kasturi, L., J. R. Eshleman, W. H. Wunner, and S. H. Shakin-Eshleman. 1995. The hydroxy amino acid in an Asn-X-Ser/Thr sequon can influence N-linked core glycosylation efficiency and the level of expression of a cell surface glycoprotein. J. Biol. Chem. 270:14756-61. Kerr, D. E., K. Plaut, A. J. Bramley, C. M. Williamson, A. J. Lax, K. Moore, A. D. Wells, and R. J. Wall. 2001. Lysostaphin expression in mammary glands confers protection against staphylococcal infection in transgenic mice. Nat. Biotechnol. 19:66-70. Kerr, D. E., and O. Wellnitz. 2003. Mammary expression of new genes to combat mastitis. J. Anim. Sci. 81:38-47. Kline, S. A., J. de la Harpe, and P. Blackburn. 1994. A colorimetric microtiter plate assay for lysostaphin using a hexaglycine substrate. Anal. Biochem. 217:329-331. Kumar, J. K. 2008. Lysostaphin: an antistaphylococcal agent. Appl. Microbial Biotechnol. 80:555-561. Lu, J. Z., T. Fujiwara, H. Komatsuzawa, M. Sugai, and J. Sakon. 2006. Cell wall-targeting domain of glycylglycine endopeptidase distinguishes among peptidoglycan cross-bridges. J. Biol. Chem. 281:549-559. Makovec, J. A., and P. L. Ruegg. 2003. Antimicrobial resistance of bacteria isolated from dairy cow milk samples submitted for bacterial culture: 8,905 samples (1994–2001). J. Am. Vet. Med. Assoc. 222:1582-1589. Miletich, J. P., and G. J. Broze Jr. 1990. Beta protein C is not glycosylated at asparagine 329. The rate of translation may influence the frequency of usage at asparagine-X-cysteine sites. J. Biol. Chem. 265:11397-11404. Moura, R. R., L. M. Melo, and V. J. F. Freitas. 2011. Production of recombinant proteins in milk of transgenic and non-transgenic goats. Braz. Arch. Biol. Technol. 54:927-938 Nilsson, I., and G. von Heijne. 1993. Determination of the distance between the oligosaccharyltransferase active site and the endoplasmic reticulum membrane. J. Biol. Chem. 268:5798-5801. Nilsson, I., G. von Heijne. 2000. Glycosylation efficiency of Asn-Xaa-Thr sequons depends both on the distance from the C terminus and on the presence of a downstream transmembrane segment. J. Biol. Chem. 275:17338-17343. Oldham, E. R., and M. J. Daley. 1991. Lysostaphin: use of a recombinant bactericidal enzyme as a mastitis therapeutic. J. Dairy Sci. 74:4175-4182. Rainard, P. 2005. Tackling mastitis in dairy cows. Nat. Biotechnol. 23:430-432. Recsei, P. A., A. D. Gruss, and R. P. Novick. 1987. Cloning, sequence, and expression of the lysostaphin gene from Staphylococcus simulans. Proc. Natl. Acad. Sci. U. S. A. 84: 1127-1131. Rogers, K. L., P. D. Fey, and M. E. Rupp. 2009. Coagulase-negative staphylococcal infections. Infect. Dis. Clin. N. Am. 23:73-98. Schindler, C. A., and V. T. Schuhardt. 1964. Lysostaphin a new bacteriolytic agent for the Staphylococcus. Proc. Natl. Acad. Sci. U. S. A. 51:414-421. Stanley, P. 1992. Glycosylation engineering. Glycobiology 2:99-107. Teale, C. J., and G. David. 1999. Antibiotic resistance in mastitis bacteria. proceedings of the British mastitis conference, Institute for Animal Health. pp. 24-29. Trayer, H. R., and C. E. Buckley. 1970. Molecular properties of lysostaphin, a bacteriolytic agent specific for Staphylococcus aureus. J. Biol. Chem. 245:4842-4846. Wall, R. J., A. M. Powell, M. J. Paape, D. E. Kerr, D. D. Bannerman, V. G. Pursel, K. D. Wells, N. Talbot, and H. W. Hawk. 2005. Genetically enhanced cows resist intramammary Staphylococcus aureus infection. Nat. Biotechnol. 23:445-451. Williamson, C. M., A. J. Bramley, and A. J. Lax. 1994. Expression of the lysostaphin gene of Staphylococcus simulans in a eukaryotic system. Appl. Environ. Microbiol. 60: 771-776. Zhang, J. X., S. F. Zhang, T. D. Wang, X. J. Guo, and R. L. Hu. 2007. Mammary gland expression of antibacterial peptide genes to inhibit bacterial pathogens causing mastitis. J. Dairy Sci. 90:5218-5225. Chapter 3 Atalay, A., T. Crook, M. Ozturk, and I. G. Yulug. 2002. Identification of genes induced by BRCA1 in breast cancer cells. Biochem. Biophys. Res. Commun. 299:839-846. Atienza, J. M., R. B. Roth, C. Rosette, K. J. Smylie, S. Kammerer, J. Rehbock, J. Ekblom, and M. F. Denissenko. 2005. Suppression of RAD21 gene expression decreases cell growth and enhances cytotoxicity of etoposide and bleomycin in human breast cancer cells. Mol. Cancer Ther. 4:361-368. Bauerschmidt, C., C. Arrichiello, S. Burdak-Rothkamm, M. Woodcock, M. A. Hill, D. L. Stevens, and K. Rothkamm. 2010. Cohesin promotes the repair of ionizing radiation-induced DNA double-strand breaks in replicated chromatin. Nucleic Acids Res. 2:477-487. Birkenbihl, R. P., and S. Subramani. 1992. Cloning and characterization of rad21 an essential gene of Schizosaccharomyces pombe involved in DNA double-strand-break repair. Nucleic Acids Res. 20:6605-6611. Birkenbihl, R. P., and S. Subramani. 1995. The rad21 gene product of Schizosaccharomyces pombe is a nuclear, cell cycle-regulated phosphoprotein. J. Biol. Chem. 270:7703-7711. Chen, F., M. Kamradt, M. Mulcahy, Y. Byun, H. Xu, M. Mckay, V. Cryns, and R. H. Lurie. 2002. Caspase proteolysis of the cohesin component RAD21 promotes apoptosis. J. Biol. Chem. 277:16775-16781. Conti, E., M. Uy, L. Leighton, G. Blobel, and J. Kuriyan. 1998. Crystallographic analysis of the recognition of a nuclear localization signal by the nuclear import factor karyopherin alpha. Cell 94:193-204. Datta, S. R., A. Brunet, and M. E. Greenberg. 1999. Cellular survival: a play in three Akts. Genes Dev. 13:2905-2927. DeMase, D., L. Zeng, C. Cera, and M. Fasullo. 2005. The Saccharomyces cerevisiae PDS1 and RAD9 checkpoint genes control different DNA double-strand break repair pathways. DNA Repair (Amst). 4:59-69. Hauf, S., I. C. Waizenegger, and J. M. Peters. 2001. Cohesin cleavage by separase required for anaphase and cytokinesis in human cells. Science 293:1320-1323. Hinz, J. M., N. A. Yamada, E. P. Salazar, R. S. Tebbs, and L. H. Thompson. 2005. Influence of double-strand-break repair pathways on radiosensitivity throughout the cell cycle in CHO cells. DNA Repair (Amst) 4:782-792. Khanna, K. K., and S. P. Jackson. 2001. DNA double-strand breaks: signaling, repair and the cancer connection. Nat. Genet. 27:247-254. Lee, J. Y., A. Hayashi-Hagihara, and T. L. Orr-Weaver. 2005. Roles and regulation of the Drosophila centromere cohesion protein MEI-S332 family. Philos. Trans. R . Soc. Lond. B. Biol. Sci. 360:543-52. Lee, J., T. Iwai, T. Yokota, and M. Yamashita. 2003. Temporally and spatially selective loss of Rec8 protein from meiotic chromosomes during mammalian meiosis. J. Cell Sci. 116:2781-2790. Lee, J. Y., and T. L. Orr-Weaver. 2001. The molecular basis of sister-chromatid cohesion. Annu. Rev. Cell Dev. Biol. 17:753-777. Martin, S. A., and T. Ouchi. 2005. BRCA1 phosphorylation regulates caspase-3 activation in UV-induced apoptosis. Cancer Res. 65:10657-10662. McKay, M. J., C. Troelstra, P. van der Spek, R. Kanaar, B. Smit, A. Hagemeijer, D. Bootsma, and J. H. Hoeijmakers. 1996. Sequence conservation of the rad21 Schizosaccharomyces pombe DNA double-strand break repair gene in human and mouse. Genomics 36:305-315. Molnar, M., J. Bahler, M. Sipiczki, and J. Kohli. 1995. The rec8 gene of Schizosaccharomyces pombe is involved in linear element formation, chromosome pairing and sister-chromatid cohesion during meiosis. Genetics 141:61-73. Panigrahi, A. K., and D. Pati. 2009. Road to the crossroads of life and death: linking sister chromatid cohesion and separation to aneuploidy, apoptosis and cancer. Crit. Rev. Oncol. Hematol. 72:181-193. Panigrphi, A. K., N. Zhang, S. K. Otta, and D. Pati. 2012. A cohesin–RAD21 interactome. Biochem. J. 442: 661-670. Pati, D., N. Zhang, and S. Plon. 2002. Linking sister chromatid cohesion and apoptosis: role of Rad21. Mol. Cell. Biol. 23:8267-8277. Porkka, K. P., T. L. Tammela, R. L. Vessella, and T. Visakorpi. 2004. RAD21 and KIAA0196 at 8q24 are amplified and overexpressed in prostate cancer. Genes Chromosomes Cancer 39:1-10. Rhodes, D. R., J. Yu, K. Shanker, N. Deshpande, R. Varambally, D. Ghosh, T. Barrette, A. Pandey, and A. M. Chinnaiyan. 2004. Large-scale meta-analysis of cancer microarray data identifies common transcriptional profiles of neoplastic transformation and progression. Proc. Natl. Acad. Sci. U. S. A. 101:9309-9314. Rothkamm, K., I. Kruger, L. H. Thompson, and M. Lobrich. 2003. Pathways of DNA double-strand break repair during the mammalian cell cycle. Mol. Cell Biol. 23:5706-5715. Scully, R., and D. M. Livingston. 2000. In search of the tumour-suppressor functions of BRCA1 and BRCA2. Nature 40:429-432. Sjogren, C., and K. Nasmyth. 2001. Sister chromatid cohesion is required for postreplicative double-strand break repair in Saccharomyces cerevisiae. Curr. Biol. 11: 991-995. Sonoda, E., T. Matsusaka, C. Morrison, P. Vagnarelli, O. Hoshi, T. Ushiki, K. Nojima, T. Fukagawa, I. C. Waizenegger, J. M. Peters, W. C. Earnshaw, and S. Takeda. 2001. Scc1/Rad21/Mcd1 is required for sister chromatid cohesion and kinetochore function in vertebrate cells. Dev. Cell 1:759-770. Strom, L., H. B. Lindroos, K. Shirahige, and C. Sjogren. 2004. Postreplicative recruitment of cohesin to double-strand breaks is required for DNA repair. Mol. Cell 16:1003-1015. Supernat, A., S. Lapińska-Szumczyk, S. Sawicki, D. Wydra, W. Biernat, and A. J. Zaczek. 2012. Deregulation of RAD21 and RUNX1 expression in endometrial cancer. Oncol. Lett. 4:727-732. Talanian, R. V., C. Quinlan, S. Trautz, M. C. Hackett, J. A. Mankovich, D. Banach, T. Ghayur, K. D. Brady, and W. W. Wong. 1997. Substrate specificities of caspase family proteases. J. Biol. Chem. 272:9677-9682. Thornberry, N., T. Rano, E. Peterson, D. Rasper, T. Timkey, M. Garcia-Calvo, V. Houtzager, P. Nordstrom, S. Roy, J. Vaillancourt, K. Chapman, and D. Nicholson. 1997. A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. J. Biol. Chem. 272:17907-17911. van’t Veer, L. J., H. Dai, M. J. van de Vijver, Y. D. He, A. A. M. Hart, M. Mao, H. L. Peterse, K. van der Kooy, M. J. Marton, A. T. Witteveen, G. J. Schreiber, R. M. Kerkhoven, C. Roberts, P. S. Linsley, R. Bernards, and S. H. Friend.. 2002. Gene expression profiling predicts clinical outcome of breast cancer. Nature 415:530-536. Xu, H., M. Yan, J. Patra, R. Natrajan, Y. Yan, S. Swagemakers, J. M. Tomaszewski, S. Verschoor, E. K. Millar, P. van der Spek, J. S. Reis-Filho, R. G. Ramsay, S. A. O'Toole ,C. M. McNeil, R. L. Sutherland, M. J. McKay, and S. B. Fox. 2011. Enhanced RAD21 cohesin expression confers poor prognosis and resistance to chemotherapy in high grade luminal, basal and HER2 breast cancers. Breast Cancer Res. 13:R9. Yamamoto, G., T, Irie, T. Aida, Y. Nagoshi, R. Tsuchiya, and T. Tachikawa. 2006. Correlation of invasion and metastasis of cancer cells, and expression of the RAD21 gene in oral squamous cell carcinoma. Virchows Arch. 448:435-441. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62952 | - |
dc.description.abstract | 隨著雌性動物懷孕,乳腺上皮細胞經歷細胞增生與分化,故其為一研究細胞生長、凋亡、移動與表現有價值重組蛋白之良好模式。然而,初級培養乳腺上皮細胞之應用受其有限生命周期之限制。本研究中,藉由表現人類端粒反轉錄酶(human telomerase reverse transcriptiase; hTERT) 與第16型人類乳突病毒E7基因建立不朽山羊乳腺上皮細胞株(CMEC-08-D)。
為確認CMEC-08-D可應用於表現具生物活性之外源性重組蛋白,將自Staphylococcus simulans選殖出之溶葡萄球菌酶(lysostaphin) 表現於CMEC-08-D,並分析其蛋白質運送路徑與醣基化位置。藉由表現帶有乳腺專一乳蛋白山羊beta-酪蛋白、乳鐵蛋白或原核訊息胜肽之成熟溶葡萄球菌酶重組蛋白於CMEC-08-D,證實兩種真核訊息胜肽皆能成功的引導重組後的成熟溶葡萄球菌酶經內質網路徑分泌至培養液中。經由突變其可能醣基化位置試驗,結果證實當溶葡萄球菌酶蛋白表現於CMEC-08-D中時,僅其第125個胺基酸-天冬醯胺酸(asparagine; Asn)的位置會發生N-鏈結醣基化作用,而第232個胺基酸-天冬醯胺的位置則否。此外,當利用點突變的方式將天冬醯胺以穀胺醯胺基酸(glutamine; Gln)取代,結果顯示突變後的N125Q與 N125Q/N232Q-溶葡萄球菌酶重組蛋白具裂解金黃色葡萄球菌能力,然而,突變的N232Q-與野生型溶葡萄球菌酶則不具裂菌活性。更進一步檢測醣基化對溶葡萄球菌酶重組蛋白與金黃色葡萄球菌結合能力之影響,結果顯示處理組間結合能力皆無差異。綜上結果顯示,第125個胺基酸上之醣基化會明顯減弱溶葡萄球菌酶之裂菌能力而非其與金黃色葡萄球菌之結合能力。 在先前的試驗中顯示,連結姊妹染色分體之黏結蛋白(cohesin) 次單位Rad21基因差異表現於小鼠乳腺的退乳時期與泌乳時期。因此,CMEC-08-D亦被用作為研究Rad21是否參與乳腺退化之研究平台。目前已知,當誘導細胞凋亡時,全長的Rad21會裂解形成N端-與C端-Rad21裂解產物。由於細胞凋亡與移動皆為乳腺退化過程之重要事件。故利用自動化即時偵測細胞行為系統(Electric Cell-substrate Impedance Sensing,ECIS)與傷口癒合分析(wound healing assay)了解Rad21與細胞移動之關係。結果顯示,外源性穩定表現全長Rad21可促進CMEC-08-D移動能力。更進一步利用紫外光(ultraviolet light)誘導細胞凋亡進而研究全長、N端-與C端-Rad21裂解產物與細胞凋亡之關係。經由細胞核染色檢測DNA片斷化情形與Akt蛋白活化情形,結果顯示外源性表現Rad21降低紫外光所誘導之細胞凋亡比例,此外,在紫外光誘導細胞凋亡後,穩定表現有Rad21的細胞株皆有較高之Akt蛋白總量。以上結果顯示,Rad21可能在乳腺退化時期維持細胞存活。 綜合以上結果,不朽化CMEC-08-D細胞株為一有價值研究模式於外源性表現具生物活性之重組蛋白。定義確切溶葡萄球菌酶之醣基化位置與醣基化對其功能之影響將有助於未來利用轉基因動物方式表現溶葡萄球菌酶之預防乳房炎之研究。此外,利用CMEC-08-D細胞株為平台亦了解Rad21可能於乳腺退化時期扮演多功能角色。 | zh_TW |
dc.description.abstract | The mammary epithelial cells undergo proliferation and differentiation during repeat pregnancy, is an ideal model for study cell growth, apoptosis, migration, and expressed valuable recombinant proteins. However, a finite life span of primary cells limits their application. Here, human telomerase reverse transcriptase (hTERT) and human papilloma virus 16 E7 genes were used to generate an immortalized caprine mammary epithelial cell line (CMEC-08-D).
To examine whether the CMEC-08-D can apply to express prokaryotic recombinant protein with biological function, an anti-staphylococcal agent lysostaphin from Staphylococcus simulans that had been used to cure Staphylococcus aureus mastitis was expressed in CMEC-08-D to study the sorting pathway and site-specific glycosylation. Recombinant lysostaphin fused with the mammary specific milk protein signal peptides of goat beta-casein, lactoferrin or prokaryotic were separately ectopic expressed in CMEC-08-D, both eukaryotic signaling peptides successfully led recombinant lysostaphins secreted into media through ER secretory pathway. Results from site-directed mutagenesis show that Asn125 but not Asn232 is the exact glycosylation site of lysostaphin expressed in CMEC-08-D. In addition, the effect of glycosylation of lysostaphin on its staphylolytic activity was identified through bacterial plate assay. The data indicated that wild type and mutated N232Q-lysostaphin (Asn232 to Gln232 substitution) lacked staphylolytic activity. In contrast, mutated N125Q (Asn125 to Gln125 substitution) and N125Q/N232Q-lysostaphin possessed staphylolytic activity. On the other hand, all mutated lysostaphin showed no change in binding ability to S. aureus. This reveals that N-glycosylation at Asn125 of lysostaphin expressed in a eukaryotic system greatly decreases lysostaphin bacteriolytic activity but does not affect its binding ability to S. aureus. In the previous study, the Rad21, one of the major cohesion subunits that hold chromatids together has highly differential expression between involution and lactation stages of the murine mammary gland. Thus, the CMEC-08-D was also used as a platform to examine whether Rad21 involved in the involution events. Rad21 is cleaved to form carboxy-terminal and amino-terminal products during apoptotic stimuli. As the cell migration and apoptosis are the important events that involved in mammary gland involution. The cell migration ability was evaluated in stable expressed exogenous Rad21 CMEC-08-D cells. The results suggested that full-length Rad21 enhances cell migration by wound healing assay and electrical cell substrate impedance-sensing system (ECIS) migration assay. To examine the effect of different truncated form Rad21 on apoptosis, ultraviolet light (UV) was used to induce cell apoptosis. The apoptosis and its pathway were evaluated by DNA fragmentation using Hoechst 33258 and Akt activation. Rad21 protected cells undergoing apoptosis induced by UV damage. Moreover, the expression of Akt was increased in Rad21 stably expressed cell lines compared with CMEC-08-D cells after UV-induced apoptosis. These data suggest that the Rad21 may maintain cell viability during mammary gland involution. Together these data, it concluded that the immortalized CMEC-08-D cells provided a valuable model for studying the biological characteristics of recombinant protein which will be expressed in mammary gland. Definition of the specific glycosylation site and the role of N-linked glycosylation on lysostaphin may provide the information to improve the efficiency of the antibacterial activity of lysostaphin in future transgenic approach. In addition, the immortalized CMEC-08-D also serving as a platform to understand Rad21 may play multifunction during mammary gland involution. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T16:16:13Z (GMT). No. of bitstreams: 1 ntu-102-D95626005-1.pdf: 24809315 bytes, checksum: 32967c2033669fa94383eb3a66c8a39c (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | Table of Contents
中文摘要..................................................i Abstract...............................................iii Introduction.............................................1 Chapter 1 Establishment of human telomerase reverse transcriptase (hTERT) and human papillomavirus (HPV)-16 E7-immortalized caprine mammary epithelial cell lines.......3 1. Literature review.....................................4 1.1 Mammary structures...................................4 1.1.1 A mammary duct.....................................4 1.1.2 Terminal end bud (TEB).............................4 1.2 Mammary gland development and differentiation........5 1.3 Application of mammary gland.........................6 1.3.1 Bioreactor...............................................6 1.3.2 Cancer research....................................6 1.4 Immortalized mammary epithelial cells................7 1.4.1 Immortalization of primary mammary epithelial cells in cultures.................................................8 1.4.2 Characterization of immortalized mammary epithelial cells in cultures........................................9 1.5 Caprine mammary gland................................9 2. Materials and Methods................................11 2.1 Immortalization of mammary epithelial cells and cell culture.................................................11 2.2 Immunoblotting for confirming HPV-16 E7 expression..12 2.3 Detection of telomerase activity....................13 2.4 Acini formation assay...............................14 3. Results..............................................16 3.1 Telomerase activity in immortalized CMEC............16 3.2 Expression of E7 protein in immortal CMEC lines.....16 3.3 Alveologenesis of immortalized CMEC-08-D............17 4. Discussion...........................................18 5. Conclusion...........................................20 6. References...........................................21 7. Figures..............................................27 Figure 1 Telomerase activity in CMEC cell lines.........27 Figure 2 Western blotting of stable E7 expression in immortalized CMEC lines.................................28 Figure 3 Immortalized CMEC-08-D cells formed alveolar structures when cultured in Matrigel....................29 Chapter 2 Application of CMEC-08-D in studying of the role of N-linked glycosylation on prokaryotic lysostaphin....30 1. Literature review....................................31 1.1 Mastitis in dairy animal............................31 1.2 Causes of mastitis..................................31 1.3 Control of mastitis.................................32 1.3.1 Progress of control mastitis......................32 1.3.2 Antimicrobial therapy.............................33 1.4 Lysostaphin (LYS)...................................34 1.5 Glycosylation of lysostaphin on prevention of Staphylococcal mastitis ................................35 1.6 N-linked glycosylation..............................36 1.7 Aim of this study...................................37 2. Materials and Methods................................38 2.1 Immortalized caprine mammaryepithelial cell line (CMEC-08-D) and cell culture..................................38 2.2 Plasmid construction................................38 2.2.1 Cloning of lysostaphin gene.......................38 2.2.2 Lysostaphin gene fused to eukaryotic expression vectors following the eukaryotic or prokaryotic signal peptide.................................................39 2.2.2.1 pcDNA4 prepro-LYS...............................39 2.2.2.2 pcDNA4 beta-casein-LYS and pcDNA4 LF-LYS........40 2.3 Site-directed mutagenesis...........................40 2.4 Transfection and collection of lysostaphin protein from culture medium..........................................42 2.5 Tunicamycin treatment...............................43 2.6 Immunoblotting......................................43 2.7 Immunocytochemistry.................................44 2.8 Bioactivity assay of lysostaphin....................45 2.9 Preparation of of Staphylococcus aureus for binding assay...................................................46 2.10 Production of concentrated recombinant lysostaphin for binding assay...........................................46 2.11 Cell wall binding assay of lysostaphin to Staphylococcus aureus...................................47 3. Results..............................................49 3.1 N-linked glycosylation inhibitor blocks the post-translational modification of eukaryotic produced lysostaphin.............................................49 3.2 N-linked glycosylation of eukaryotic produced lysostaphin loses its staphylolytic activity............49 3.3 Generation of signal peptide fused wild type and N-linked glycosylation site mutated lysostaphin constructs ........................................................50 3.4 The sorting of various recombinant lysostaphins in CMEC-08-D....................................................52 3.5 Recombinant lysostaphins with eukaryotic signal peptides secreted by CMEC-08-D...................................53 3.6 Asn125 N-linked glycosylation of lysostaphin decreased lysostaphin catalytic activity but not the ability to bind to S aureus.............................................54 4. Discussion...........................................57 4.1 btea-Casein and lactoferrin signal peptides direct secretion of recombinant lysostaphin in CMEC-08-D.......57 4.2 Amino acid Asn125 residue is the specific N-linked glycosylation site on eukaryotically produced recombinant lysostaphin.............................................59 4.3 Glycosylation at Asn125 residue of recombinant lysostaphin critically decreases lysostaphin lytic activity but not the binding ability to S aureus.................60 5. Conclusion...........................................63 6. References...........................................64 7. Table................................................71 Table1 Template and primer list for generation of various mutated lysostaphins....................................71 8. Figures..............................................72 Figure 1 Tunicamycin inhibits N-linked glycosylation of lysostaphin in CMEC-08-D cells..........................72 Figure 2 N-linked glycosylation lysostaphin from CMEC-08-D cells inhibits staphylolytic activity to S. aureus......74 Figure 3 Schematic representation of the constructs and sequences of various recombinant lysostaphins...........75 Figure 4 The sorting patterns of various mutated recombinant lysostaphins leading with beta-casein or lactoferrin signal peptides in CMEC-08-D were analyzed by immunoflurorescent staining................................................77 Figure 5 The analysis polypeptide mass of N-linked glycosylation site mutated and wild type of secreted lysostaphin from CMEC-08-D cells by Western blotting....80 Figure 6 Asn125 N-linked glycosylation site of lysostaphin is responsible for the staphylolytic activity of lysostaphin produced by CMEC-08-D...................................82 Figure 7 Binding ability of N-linked glycosylation and mutanted lysostaphin to Staphylococcus aureus...........84 9. Appendix.............................................86 Supplemental 1 The sorting patterns of various mutated recombinant lysostaphins leading with beta-casein or lactoferrin signal peptides in CMEC-08-D were analyzed by immunoflurorescent staining.............................86 Chapter 3 Use of CMEC-08-D to study the effect of Rad21 on the events of mammary involution........................89 1. Literature review....................................90 1.1 Rad21...............................................90 1.2 Rad21 Protein structure.............................91 1.3 Functional studies of Rad21.........................92 1.3.1 Rad21 in mitosis..................................92 1.3.2 Rad21 in DNA double-strand-break repair...........93 1.3.3 Rad21 in apoptosis................................94 1.3.4 Rad21 in cancer...................................96 2. Materials and Methods................................98 2.1 Plasmids............................................98 2.1.1 Cloning of Rad21 gene.............................98 2.1.2 Expression plasmid construction...................99 2.2 Generation of Rad21-expressing CMEC-08-D...........100 2.3 Immunoblotting.....................................101 2.4 Immuocytochemisty..................................102 2.5 UV-induced apoptosis...............................103 2.6 Wound healing assay................................104 2.7 Electric cell substrate impedance sensing (ECIS) wound healing assay..........................................104 2.8 Statistical analysis...............................105 3. Results.............................................106 3.1 Establishment of stable cell line with normal and different truncated form Rad21.........................106 3.2 Effect of Rad21 on the motility of CMEC-08-D.......107 3.3 Rad21 protecting cells from UV-induced apoptosis...108 4. Discussion..........................................111 4.1 Subcellular localization and molecular mass of various Rad21..................................................111 4.2 The effect of Rad21 on cellular motility...........113 4.3 The effect of Rad21 on anti-apoptosis..............114 5. Conclusion..........................................117 6. References..........................................118 7. Figures.............................................124 Figure 1 Immunofluorescence staining of CMEC-08-D cells stably expressing exogenous different truncated form of Rad21..................................................124 Figure 2 Western blot analysis of various Rad21 molecular mass in Rad21 stably expressed cell line...............125 Figure 3 Full-length Rad21 promote cell migration as determined by electric cell-substrate impedance sensing (ECIS) wound healing assay.............................126 Figure 4 Full-length Rad21 enhanced cell migration ability in wound healing assay ................................127 Figure 5 Evidence of various Rad21 protects cells from apoptosis after UV-induced apoptosis...................129 Figure 6 Rad21 increased total Akt expression after UV radiation treatment of CMEC-08-D.......................131 Summary and future work................................133 | |
dc.language.iso | en | |
dc.title | 應用不朽化山羊乳腺上皮細胞研究醣基化對原核溶葡萄球菌酶之影響與真核Rad21基因於細胞存活路徑之角色 | zh_TW |
dc.title | Application of immortalized caprine mammary epithelial cells for investigating the glycosylation effect on prokaryotic lysostaphin and the role of eukaryotic Rad21 on survival pathway | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-1 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 姜延年(Yan-Nian Jiang) | |
dc.contributor.oralexamcommittee | 徐濟泰,黃木秋,陳銘正,楊瀅臻 | |
dc.subject.keyword | 山羊,乳腺上皮細胞,溶葡萄球菌酶,N-鏈結醣基化作用,Rad21,細胞凋亡, | zh_TW |
dc.subject.keyword | caprine,mammary epithelial cells,lysostaphin,N-glycosylation,Rad21,apoptosis, | en |
dc.relation.page | 134 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-02-05 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 動物科學技術學研究所 | zh_TW |
顯示於系所單位: | 動物科學技術學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-102-1.pdf 目前未授權公開取用 | 24.23 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。