請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71247
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林欽塘(Chin-Tarng Lin) | |
dc.contributor.author | Ting-Ying Chen | en |
dc.contributor.author | 陳亭螢 | zh_TW |
dc.date.accessioned | 2021-06-17T05:00:47Z | - |
dc.date.available | 2028-07-25 | |
dc.date.copyright | 2018-08-30 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-07-25 | |
dc.identifier.citation | 1. Ei-Naggar, A., et al., WHO Classification of Head and Neck Tumours. 2017, Lyon: IAPR press.
2. Torre, L.A., et al., Global cancer statistics, 2012. CA: a cancer journal for clinicians, 2015. 65(2): p. 87-108. 3. Parkin, D.M., et al., Estimating the world cancer burden: Globocan 2000. International journal of cancer, 2001. 94(2): p. 153-156. 4. Shanmugaratnam, K., L.H. Sobin, and W.H. Organization, Histological typing of upper respiratory tract tumours/K. Shanmugaratnam, in collaboration with LH Sobin and pathologists in 10 countries, in Histological typing of upper respiratory tract tumours/K. Shanmugaratnam, in collaboration with LH Sobin and pathologists in 10 countries. 1978. 5. Müller, E. and E. Beleites, The basaloid squamous cell carcinoma of the nasopharynx. Rhinology, 2000. 38(4): p. 208-211. 6. Banks, E.R., et al., Basaloid squamous cell carcinoma of the head and neck. A clinicopathologic and immunohistochemical study of 40 cases. The American journal of surgical pathology, 1992. 16(10): p. 939-946. 7. Hanley, A.J., B.C. Choi, and E.J. Holowaty, Cancer mortality among Chinese migrants: a review. International journal of epidemiology, 1995. 24(2): p. 255-265. 8. Ren, Z.-F., et al., Effect of family history of cancers and environmental factors on risk of nasopharyngeal carcinoma in Guangdong, China. Cancer epidemiology, 2010. 34(4): p. 419-424. 9. Yu, M.C., T.B. Huang, and B.E. Henderson, Diet and nasopharyngeal carcinoma: A case‐control study in Guangzhou, China. International journal of cancer, 1989. 43(6): p. 1077-1082. 10. Ward, M.H., et al., Dietary exposure to nitrite and nitrosamines and risk of nasopharyngeal carcinoma in Taiwan. International journal of cancer, 2000. 86(5): p. 603-609. 11. Cheng, Y.-J., et al., Cigarette smoking, alcohol consumption and risk of nasopharyngeal carcinoma in Taiwan. Cancer Causes & Control, 1999. 10(3): p. 201-207. 12. Armstrong, R.W., et al., Nasopharyngeal carcinoma in Malaysian Chinese: occupational exposures to particles, formaldehyde and heat. International journal of epidemiology, 2000. 29(6): p. 991-998. 13. Miller, G., Epstein-Barr virus; biology, pathogenesis, and medicalaspect. Virology, 1990. 14. Vetsika, E.-K. and M. Callan, Infectious mononucleosis and Epstein-Barr virus. Expert reviews in molecular medicine, 2004. 6(23): p. 1-16. 15. Raab‐Traub, N., et al., The differentiated form of nasopharyngeal carcinoma contains epstein‐barr virus DNA. International journal of cancer, 1987. 39(1): p. 25-29. 16. Chan, K.A. and Y.D. Lo. Circulating EBV DNA as a tumor marker for nasopharyngeal carcinoma. in Seminars in cancer biology. 2002. Elsevier. 17. Maxwell, J.H., et al., HPV‐positive/p16‐positive/EBV‐negative nasopharyngeal carcinoma in white North Americans. Head & neck, 2010. 32(5): p. 562-567. 18. Hildesheim, A. and C.-P. Wang. Genetic predisposition factors and nasopharyngeal carcinoma risk: a review of epidemiological association studies, 2000–2011: Rosetta Stone for NPC: genetics, viral infection, and other environmental factors. in Seminars in cancer biology. 2012. Elsevier. 19. Simons, M., et al., Immunogenetic aspects of nasopharyngeal carcinoma: I. Differences in HL‐A antigen profiles between patients and control groups. International journal of cancer, 1974. 13(1): p. 122-134. 20. Tang, M., et al., Haplotype-dependent HLA susceptibility to nasopharyngeal carcinoma in a Southern Chinese population. Genes and immunity, 2010. 11(4): p. 334. 21. Kelly, J.Y., et al., Association of human leukocyte antigens with nasopharyngeal carcinoma in high-risk multiplex families in Taiwan. Human immunology, 2009. 70(11): p. 910-914. 22. Hildesheim, A., et al., Association of HLA class I and II alleles and extended haplotypes with nasopharyngeal carcinoma in Taiwan. Journal of the National Cancer Institute, 2002. 94(23): p. 1780-1789. 23. Karanikiotis, C., et al., HLA class II alleles and the presence of circulating Epstein-Barr virus DNA in Greek patients with nasopharyngeal carcinoma. Strahlentherapie und Onkologie, 2008. 184(6): p. 325-331. 24. Li, X., et al., Associations between HLA class I alleles and the prevalence of nasopharyngeal carcinoma (NPC) among Tunisians. Journal of translational medicine, 2007. 5(1): p. 22. 25. Jalbout, M., et al., Polymorphism of the stress protein HSP70-2 gene is associated with the susceptibility to the nasopharyngeal carcinoma. Cancer letters, 2003. 193(1): p. 75-81. 26. Douik, H., et al., Association of MICA-129 polymorphism withnasopharyngeal cancer risk in a Tunisian population. Human immunology, 2009. 70(1): p. 45-48. 27. Yang, Z.H., et al., Association of IL‐1 polymorphisms and IL‐1 serum levels with susceptibility to nasopharyngeal carcinoma. Molecular carcinogenesis, 2011. 50(3): p. 208-214. 28. Wei, Y.-S., et al., Association of the interleukin-2 polymorphisms with interleukin-2 serum levels and risk of nasopharyngeal carcinoma. DNA and cell biology, 2010. 29(7): p. 363-368. 29. Wei, Y.-S., et al., Single nucleotide polymorphism and haplotype association of the interleukin-8 gene with nasopharyngeal carcinoma. Clinical Immunology, 2007. 125(3): p. 309-317. 30. Wei, Y.S., et al., Interleukin‐10 gene promoter polymorphisms and the risk of nasopharyngeal carcinoma. HLA, 2007. 70(1): p. 12-17. 31. Gao, L.-B., et al., Genetic polymorphism of interleukin-16 and risk of nasopharyngeal carcinoma. Clinica Chimica Acta, 2009. 409(1-2): p. 132-135. 32. Nong, L.-G., et al., Interleukin-18 gene promoter polymorphism and the risk of nasopharyngeal carcinoma in a Chinese population. DNA and cell biology, 2009. 28(10): p. 507-513. 33. Farhat, K., et al., Interleukin‐10 and interferon‐gamma gene polymorphisms in patients with nasopharyngeal carcinoma. International journal of immunogenetics, 2008. 35(3): p. 197-205. 34. Wei, Y.-S., et al., Association of transforming growth factor-β1 gene polymorphisms with genetic susceptibility to nasopharyngeal carcinoma. Clinica Chimica Acta, 2007. 380(1-2): p. 165-169. 35. He, J.-F., et al., Genetic polymorphisms of TLR3 are associated with Nasopharyngeal carcinoma risk in Cantonese population. BMC cancer, 2007. 7(1): p. 194. 36. Song, C., et al., Functional variant in the 3’-untranslated region of Toll-like receptor 4 is associated with nasopharyngeal carcinoma risk. Cancer biology & therapy, 2006. 5(10): p. 1285-1291. 37. Zhou, X.-X., et al., Sequence variants in toll-like receptor 10 are associated with nasopharyngeal carcinoma risk. Cancer Epidemiology and Prevention Biomarkers, 2006. 15(5): p. 862-866. 38. Xu, Y.-F., et al., Sequencing of DC-SIGN promoter indicates an association between promoter variation and risk of nasopharyngeal carcinoma in cantonese. BMC medical genetics, 2010. 11(1): p. 161. 39. Kovacic, M.B., et al., Variation of the killer cell immunoglobulin-like receptors and HLA-C genes in nasopharyngeal carcinoma. Cancer Epidemiology and Prevention Biomarkers, 2005. 14(11): p. 2673-2677. 40. Kongruttanachok, N., et al., Cytochrome P450 2E1 polymorphismand nasopharyngeal carcinoma development in Thailand: a correlative study. BMC cancer, 2001. 1(1): p. 4. 41. Cho, E.-Y., et al., Nasopharyngeal carcinoma and genetic polymorphisms of DNA repair enzymes XRCC1 and hOGG1. Cancer Epidemiology and Prevention Biomarkers, 2003. 12(10): p. 1100-1104. 42. Yang, Z.-H., et al., Genetic polymorphisms of the DNA repair gene and risk of nasopharyngeal carcinoma. DNA and cell biology, 2007. 26(7): p. 491-496. 43. Yang, Z.-H., et al., The xeroderma pigmentosum group C gene polymorphisms and genetic susceptibility of nasopharyngeal carcinoma. Acta Oncologica, 2008. 47(3): p. 379-384. 44. Yang, Z.H., et al., Association of ERCC1 polymorphisms and susceptibility to nasopharyngeal carcinoma. Molecular carcinogenesis, 2009. 48(3): p. 196-201. 45. Zhou, G., et al., MDM2 promoter SNP309 is associated with risk of occurrence and advanced lymph node metastasis of nasopharyngeal carcinoma in Chinese population. Clinical cancer research, 2007. 13(9): p. 2627-2633. 46. Xiao, M., et al., Genetic polymorphisms of MDM2 and TP53 genes are associated with risk of nasopharyngeal carcinoma in a Chinese population. BMC cancer, 2010. 10(1): p. 147. 47. Ma, F., et al., Functional polymorphism-31C/G in the promoter of BIRC5 gene and risk of nasopharyngeal carcinoma among chinese. PLoS One, 2011. 6(2): p. e16748. 48. Nasr, H.B., et al., A single nucleotide polymorphism in the E-cadherin gene promoter− 160 C/A is associated with risk of nasopharyngeal cancer. Clinica Chimica Acta, 2010. 411(17-18): p. 1253-1257. 49. Nasr, H.B., et al., Matrix metalloproteinase-1 (-1607) 1G/2G and-9 (–1562) C/T promoter polymorphisms: Susceptibility and prognostic implications in nasopharyngeal carcinomas. Clinica chimica acta, 2007. 384(1-2): p. 57-63. 50. Shao, J.-Y., et al., A single nucleotide polymorphism in the matrix metalloproteinase 2 promoter is closely associated with high risk of nasopharyngeal carcinoma in Cantonese from southern China. Chinese journal of cancer, 2011. 30(9): p. 620. 51. Nasr, H.B., et al., PTGS2 (COX-2)− 765 G> C functional promoter polymorphism and its association with risk and lymph node metastasis in nasopharyngeal carcinoma. Molecular biology reports, 2009. 36(1): p. 193-200. 52. Wang, T., et al., Polymorphism of VEGF-2578C/A associated withthe risk and aggressiveness of nasopharyngeal carcinoma in a Chinese population. Molecular biology reports, 2010. 37(1): p. 59-65. 53. FitzGerald, P.C., et al., Clustering of DNA sequences in human promoters. Genome research, 2004. 14(8): p. 1562-1574. 54. Virbasius, C.-m.A., J.V. Virbasius, and R.C. Scarpulla, NRF-1, an activator involved in nuclear-mitochondrial interactions, utilizes a new DNA-binding domain conserved in a family of developmental regulators. Genes & development, 1993. 7(12a): p. 2431-2445. 55. Roy, D. and R. Tamuli, NRF1 (nuclear respiratory factor 1). 2009. 56. Gugneja, S. and R.C. Scarpulla, Serine phosphorylation within a concise amino-terminal domain in nuclear respiratory factor 1 enhances DNA binding. Journal of Biological Chemistry, 1997. 272(30): p. 18732-18739. 57. Herzig, R.P., S. Scacco, and R.C. Scarpulla, Sequential serum-dependent activation of CREB and NRF-1 leads to enhanced mitochondrial respiration through the induction of cytochrome c. Journal of Biological Chemistry, 2000. 275(17): p. 13134-13141. 58. Evans, M.J. and R. Scarpulla, Interaction of nuclear factors with multiple sites in the somatic cytochrome c promoter. Characterization of upstream NRF-1, ATF, and intron Sp1 recognition sequences. Journal of Biological Chemistry, 1989. 264(24): p. 14361-14368. 59. Okoh, V., A. Deoraj, and D. Roy, Estrogen-induced reactive oxygen species-mediated signalings contribute to breast cancer. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, 2011. 1815(1): p. 115-133. 60. Piantadosi, C.A. and H.B. Suliman, Mitochondrial transcription factor A induction by redox activation of nuclear respiratory factor 1. Journal of Biological Chemistry, 2006. 281(1): p. 324-333. 61. Scarpulla, R.C., Transcriptional paradigms in mammalian mitochondrial biogenesis and function. Physiological reviews, 2008. 88(2): p. 611-638. 62. Kelly, D.P. and R.C. Scarpulla, Transcriptional regulatory circuits controlling mitochondrial biogenesis and function. Genes & development, 2004. 18(4): p. 357-368. 63. Clayton, D.A., Transcription and replication of animal mitochondrial DNAs, in International review of cytology. 1992, Elsevier. p. 217-232. 64. Fisher, R.P. and D.A. Clayton, Purification and characterization of human mitochondrial transcription factor 1. Molecular and cellular biology, 1988. 8(8): p. 3496-3509.65. Gleyzer, N., K. Vercauteren, and R.C. Scarpulla, Control of mitochondrial transcription specificity factors (TFB1M and TFB2M) by nuclear respiratory factors (NRF-1 and NRF-2) and PGC-1 family coactivators. Molecular and cellular biology, 2005. 25(4): p. 1354-1366. 66. Evans, M.J. and R.C. Scarpulla, NRF-1: a trans-activator of nuclear-encoded respiratory genes in animal cells. Genes & development, 1990. 4(6): p. 1023-1034. 67. May, B.K., et al., Molecular regulation of heme biosynthesis in higher vertebrates, in Progress in nucleic acid research and molecular biology. 1995, Elsevier. p. 1-51. 68. Kappas, A., The porphyrias. The metabolic basis of inherited disease, 1989: p. 1305-1365. 69. Braidotti, G., I. Borthwick, and B. May, Identification of regulatory sequences in the gene for 5-aminolevulinate synthase from rat. Journal of Biological Chemistry, 1993. 268(2): p. 1109-1117. 70. Aizencang, G.I., et al., Uroporphyrinogen III Synthase an alternative promoter controls erythroid-specific expression in the murine gene. Journal of Biological Chemistry, 2000. 275(4): p. 2295-2304. 71. Truscott, K.N., K. Brandner, and N. Pfanner, Mechanisms of protein import into mitochondria. Current Biology, 2003. 13(8): p. R326-R337. 72. Blesa, J.R., et al., NRF-2 transcription factor is required for human TOMM20 gene expression. Gene, 2007. 391(1): p. 198-208. 73. Takahashi, Y., et al., Characterization and identification of promoter elements in the mouse COX17 gene. Biochimica et Biophysica Acta (BBA)-Gene Structure and Expression, 2002. 1574(3): p. 359-364. 74. Piantadosi, C.A. and H.B. Suliman, Redox regulation of mitochondrial biogenesis. Free Radical Biology and Medicine, 2012. 53(11): p. 2043-2053. 75. Dominy Jr, J.E., et al., Nutrient-dependent regulation of PGC-1α's acetylation state and metabolic function through the enzymatic activities of Sirt1/GCN5. Biochimica et biophysica acta (BBA)-proteins and proteomics, 2010. 1804(8): p. 1676-1683. 76. Bergeron, R., et al., Chronic activation of AMP kinase results in NRF-1 activation and mitochondrial biogenesis. American Journal of Physiology-Endocrinology And Metabolism, 2001. 281(6): p. E1340-E1346. 77. Reznick, R.M. and G.I. Shulman, The role of AMP‐activated protein kinase in mitochondrial biogenesis. The Journal of physiology, 2006. 574(1): p. 33-39. 78. Nisoli, E., et al., Mitochondrial biogenesis in mammals: the role ofendogenous nitric oxide. Science, 2003. 299(5608): p. 896-899. 79. Niida, A., et al., Integrative bioinformatics analysis of transcriptional regulatory programs in breast cancer cells. BMC bioinformatics, 2008. 9(1): p. 404. 80. Radde, B.N., et al., Nuclear respiratory factor-1 and bioenergetics in tamoxifen-resistant breast cancer cells. Experimental cell research, 2016. 347(1): p. 222-231. 81. Blesa, J.R., et al., NRF-1 is the major transcription factor regulating the expression of the human TOMM34 gene. Biochemistry and Cell Biology, 2008. 86(1): p. 46-56. 82. Patti, M.E., et al., Coordinated reduction of genes of oxidative metabolism in humans with insulin resistance and diabetes: Potential role of PGC1 and NRF1. Proceedings of the national academy of sciences, 2003. 100(14): p. 8466-8471. 83. Liu, Y., et al., Genetic variation and association analyses of the nuclear respiratory factor 1 (nRF1) gene in Chinese patients with type 2 diabetes. Diabetes, 2008. 57(3): p. 777-782. 84. Hashad, D., I. Elgohry, and F. Dwedar, Nuclear Respiratory Factor-1 (NRF-1) Gene Expression in Chronic Kidney Disease Patients Undergoing Hemodialysis and Mitochondrial Oxidative Dysregulation. Clinical laboratory, 2016. 62(11): p. 2149-2154. 85. Lin, C., et al., Establishment and characterization of two nasopharyngeal carcinoma cell lines. Laboratory investigation; a journal of technical methods and pathology, 1990. 62(6): p. 713-724. 86. Lin, C.-T., et al., Characterization of seven newly established nasopharyngeal carcinoma cell lines. Laboratory investigation; a journal of technical methods and pathology, 1993. 68(6): p. 716-727. 87. Witkiewicz, A.K., et al., Molecular profiling of a lethal tumor microenvironment, as defined by stromal caveolin-1 status in breast cancers. Cell cycle, 2011. 10(11): p. 1794-1809. 88. Huang, D.Y., et al., Transcription factor SOX‐5 enhances nasopharyngeal carcinoma progression by down‐regulating SPARC gene expression. The Journal of pathology, 2008. 214(4): p. 445-455. 89. Ahmad, A. and S. Stefani, Distant metastases of nasopharyngeal carcinoma: a study of 256 male patients. Journal of surgical oncology, 1986. 33(3): p. 194-197. 90. Jezierska-Drutel, A., S.A. Rosenzweig, and C.A. Neumann, Role of oxidative stress and the microenvironment in breast cancer development and progression, in Advances in cancer research. 2013, Elsevier. p. 107-125. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71247 | - |
dc.description.abstract | 鼻咽癌為一種上皮細胞癌,好發於中國南方、東南亞、台灣及北非。目前其確實之致病機轉仍不是很清楚,但可能與EB病毒 ( Epstein-Barr virus ) 之感染、環境因子及遺傳因素有關。本論文研究的主要目的是探討鼻咽癌的發生與基因的關係。我們運用cDNA微陣列分析 ( cDNA Microarray Analysis ) 的方法比較鼻咽癌細胞株以及鼻腔內膜上表皮細胞之基因表現的差異,選出了11個在兩者之間有差異表現的基因,然後再經由即時定量聚合酶連鎖反應 ( Quantitative RT-PCR )的分析而發現FGFR1基因及SPARC基因在鼻咽癌細胞中的表現明顯減少;接著我們進一步觀察與FGFR1基因及SPARC基因有關的轉錄因子NRF1 ( Nuclear respiratory factor 1 ),再經由即時定量聚合酶連鎖反應 ( Quantitative RT-PCR ) 和西方點墨法 ( Western blotting ) 的確認分析,及細胞免疫染色法的觀察後,我們發現NRF1基因在鼻咽癌細胞的細胞核有高表現的現象。為了更進一步了解NRF1基因與鼻咽癌的關係,我們利用CRISPR / Cas9的技術將本實驗室所建立的TW01人類鼻咽癌細胞株的NRF1基因剔除,使其NRF1表現量下降,藉此觀察此基因在鼻咽癌病理機制上所扮演的角色及功能。我們首先利用西方點墨法 ( Western blotting ) 確認NRF1基因剔除之鼻咽癌細胞之NRF1蛋白表現量是否有顯著的下降,接著挑選出38株single cell lines,再從中挑選剔除效果最好的細胞株來進行接下來的實驗。我們發現剔除NRF1的鼻咽癌細胞的爬行速度及生長速度均較原本的鼻咽癌細胞株緩慢,且以雙氧水增加細胞氧化壓力的實驗發現剔除NRF1的鼻咽癌細胞較容易死亡,接著於氧氣消耗速率及細胞糖解作用的實驗都發現剔除NRF1的鼻咽癌細胞在此兩種代謝中都有下降的情形。在動物實驗的部分,我們發現打入剔除NRF1鼻咽癌細胞的Nude小鼠其腫瘤生長情形較正常鼻咽癌細胞的Nude小鼠緩慢。由體外的細胞實驗及動物實驗結果,我們推論NRF1在鼻咽癌的形成扮演一種致癌基因的角色。 | zh_TW |
dc.description.abstract | Nasopharyngeal carcinoma ( NPC ) is arising from nasopharyngeal epithelium. Endemic regions of NPC are shown in southern China, southeast Asia, northern Africa and Taiwan. The specific etiology is not clear now, but some evidences show that NPC is related to environmental factors, heredity, and Epstein-Barr virus infection . The purpose of this research was to find out the genes associated with NPC pathogenesis. Previously we compared the mRNA expression between NPC cell lines and normal nasal mucosal epithelial cells by cDNA microarray analysis, and found that 11 genes showed significantly increased or decreased expression. Then we used quantitative RT-PCR for further confirmation, and found that FGFR1 gene and SPARK gene were significantly decreased in NPC cell lines. In addition, we also found that the transcription factor NRF1 ( Nuclear respiratory factor 1 ) could regulate the FGFR1 gene and SPARK gene expression. When we compared NRF1 gene expression between NPC cell lines and normal nasal mucosal epithelial cells by quantitative RT-PCR, Western blotting and immunocytochemistry, we found that NRF1 gene was significantly increased expression in NPC cell lines. In order to understand the role of NRF1 expression in nasopharyngeal carcinoma, we used CRISPR/Cas9 to knockout NRF1 gene in NPC TW01 cell line which was established from our lab. After knockout of NRF1, we used Western blotting to confirm the NRF1 expression in NPC cells, and found that the NRF1 protein was significantly decreased. Then we produced 38 single stable cell lines, and chose the best gene knockout clone for the following experiments. We found that NRF1 knockout could decrease cell proliferation, migration, and invasion activities of NPC cells. When we added hydrogen peroxide into medium to increase oxidative stress, we found that NPC cells which expressed lower NRF1 had worse viability. As for metabolism, we found that both of mitochondrial respiration ability and glycolysis ability were downregulated in knockout of NRF1 cells. The in vivo animal experiment by injection of NRF1 knockout cells also revealed that the tumors growth was decreased, which confirmed the results of in vitro assay. According to in vitro assay and xenograft experiment, we suggest that NRF1 plays a role as an oncogene in nasopharyngeal carcinoma pathogenesis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T05:00:47Z (GMT). No. of bitstreams: 1 ntu-107-R05444004-1.pdf: 5624044 bytes, checksum: 59d4ea773b3963acdb01114083dbdfe5 (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 口試委員會審定書 ........................................................................................................... I
致謝 ................................................................................................................................. II 中文摘要 ........................................................................................................................ III Abstract ......................................................................................................................... IV List of figures ................................................................................................................ IX List of tables .................................................................................................................... X List of abbreviations ..................................................................................................... XI Introduction .................................................................................................................... 1 1.1 Nasopharyngeal carcinoma ( NPC ) and Epidemiology ................................ 1 1.2 Nasopharyngeal carcinoma ( NPC ) WHO classification .............................. 2 1.3 Etiology of nasopharyngeal carcinoma ( NPC )............................................. 3 (1) Environmental factors ........................................................................... 3 (2) Virus infection ....................................................................................... 4 (3) Genetic susceptibility ............................................................................ 4 a. Immune-related genes .................................................................... 5 b. DNA-repair genes .......................................................................... 6 c. Other genes related to NPC ........................................................... 6 1.4 Nuclear respiratory factor 1 ( NRF1 ) ............................................................ 7 1.5 Mitochondria and Nuclear respiratory factor 1 ( NRF1 )............................... 8 (1) NRF1 and mtDNA transcription ........................................................... 9 (2) NRF1 and heme biosynthesis .............................................................. 10 (3) NRF1 and mitochondrial proteins import and assembly ..................... 10 (4) NRF1 and redox regulation of mitochondrial biogenesis ................... 10 1.6 Nuclear respiratory factor 1 ( NRF1 ) and Disease ...................................... 11 (1) NRF1 and breast cancer ...................................................................... 11 (2) NRF1 and colorectal cancer ................................................................ 12 (3) NRF1 and type 2 diabetes mellitus ( DM ) ......................................... 12 (4) NRF1 and chronic kidney disease ....................................................... 12 Materials and Methods ................................................................................................ 13 2.1 Cell lines ....................................................................................................... 13 2.2 Extraction of RNA, preparation of cDNA, and quantitative reverse transcription polymerase chain reaction ( qRT-PCR ) .......................................... 13 2.3 Protein extraction and Western blotting ....................................................... 15 2.4 Immunocytochemistry ( ICC ) ..................................................................... 16 2.5 Immunohistochemistry ( IHC ) .................................................................... 17 2.6 Hematoxylin and eosin stain ( H&E stain ).................................................. 18 2.7 Establishment of knockout NRF1 NPC cell line .......................................... 18 2.8 Scratch wound healing assay........................................................................ 19 2.9 MTT assay .................................................................................................... 20 2.10 Invasion assay ............................................................................................ 20 2.11 Cell Mitochondria ( Mito ) Stress Test....................................................... 20 2.12 Glycolysis Stress Test ................................................................................ 21 2.13 In vivo assay of xenograft growth .............................................................. 22 2.14 Statistical analysis ......................................................................................... 23 Results ............................................................................................................................ 24 3.1 NRF1 expression in NNM and NPC cell lines ............................................. 24 3.2 NRF1 protein expression in NPC biopsy specimens ................................... 25 (1) NRF1 protein expression in non-keratinizing squamous cell carcinoma..................................................................................................................... 25 (2) NRF1 protein expression in keratinizing squamous cell carcinoma ... 26 3.3 Establishment of knockout NRF1 NPC cell lines ........................................ 26 3.4 Functional analysis of NRF1 gene expression ............................................. 28 (1) Knockout of NRF1 reduced the proliferation ability in NPC cells ..... 28 (2) Knockout of NRF1 reduced the migration ability in NPC cells ......... 28 (3) Knockout of NRF1 reduced the invasion activity in NPC cells .......... 29 (4) Knockout of NRF1 lead to cell death under ROS stress in NPC cells 29 (5) Knockout of NRF1 reduced the mitochondrial respiration ability in NPC cells ..................................................................................................... 29 (6) Knockout of NRF1 reduced the glycolysis ability in NPC cells ......... 30 3.5 The growth-suppressing effect of NRF1 in NPC xenograft Nude mice ...... 30 Discussion ...................................................................................................................... 32 Figures ........................................................................................................................... 36 Tables ............................................................................................................................. 56 References..................................................................................................................... 65 | |
dc.language.iso | en | |
dc.title | Nrf1基因在鼻咽癌之功能分析 | zh_TW |
dc.title | Functional Analysis of Nrf1 Gene in
Nasopharyngeal Carcinoma | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 吳漢忠,張久瑗(Jeou-Yuan Chen),林中梧(Chung-Wu Lin),楊雅倩 | |
dc.subject.keyword | 鼻咽癌,NRF1,CRISPR / Cas9, | zh_TW |
dc.subject.keyword | NPC,NRF1,CRISPR/ Cas9,Nude mice, | en |
dc.relation.page | 71 | |
dc.identifier.doi | 10.6342/NTU201801833 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-07-25 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 病理學研究所 | zh_TW |
顯示於系所單位: | 病理學科所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-107-1.pdf 目前未授權公開取用 | 5.49 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。