請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66666
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林欽塘 | |
dc.contributor.author | Cheng-Der Wu | en |
dc.contributor.author | 吳承德 | zh_TW |
dc.date.accessioned | 2021-06-17T00:49:55Z | - |
dc.date.available | 2017-03-02 | |
dc.date.copyright | 2012-03-02 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-11-29 | |
dc.identifier.citation | 1. Kwak PB, Iwasaki S, Tomari Y: The microRNA pathway and cancer. Cancer Sci 2010, 101:2309-2315.
2. Li M, Li J, Ding X, He M, Cheng SY: microRNA and cancer. AAPS J 2010, 12:309-317. 3. Lima RT, Busacca S, Almeida GM, Gaudino G, Fennell DA, Vasconcelos MH: MicroRNA regulation of core apoptosis pathways in cancer. Eur J Cancer 2011, 47:163-174. 4. Zimmerman AL, Wu S: MicroRNAs, cancer and cancer stem cells. Cancer Lett 2011, 300:10-19. 5. Huang Y, Shen XJ, Zou Q, Wang SP, Tang SM, Zhang GZ: Biological functions of microRNAs: a review. J Physiol Biochem 2011, 67:129-139. 6. Lee RC, Feinbaum RL, Ambros V: The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 1993, 75:843-854. 7. Lagos-Quintana M, Rauhut R, Meyer J, Borkhardt A, Tuschl T: New microRNAs from mouse and human. RNA 2003, 9:175-179. 8. Jost D, Nowojewski A, Levine E: Small RNA biology is systems biology. BMB Rep 2011, 44:11-21. 9. Gottesman S, Storz G: Bacterial Small RNA Regulators: Versatile Roles and Rapidly Evolving Variations. Cold Spring Harb Perspect Biol 2010. 10. Wu TH, Chang IY, Chu LC, Huang HC, Ng WV: Modularity of Escherichia coli sRNA regulation revealed by sRNA-target and protein network analysis. BMC Bioinformatics 2010, 11 Suppl 7:S11. 11. Caron MP, Lafontaine DA, Masse E: Small RNA-mediated regulation at the level of transcript stability. RNA Biol 2010, 7:140-144. 12. Du T, Zamore PD: microPrimer: the biogenesis and function of microRNA. Development 2005, 132:4645-4652. 13. Gregory RI, Chendrimada TP, Cooch N, Shiekhattar R: Human RISC couples microRNA biogenesis and posttranscriptional gene silencing. Cell 2005, 123:631-640. 14. Farazi TA, Spitzer JI, Morozov P, Tuschl T: miRNAs in human cancer. J Pathol 2011, 223:102-115. 15. Kozomara A, Griffiths-Jones S: miRBase: integrating microRNA annotation and deep-sequencing data. Nucleic Acids Res 2011, 39:D152-157. 16. Ambros V, Bartel B, Bartel DP, Burge CB, Carrington JC, Chen X, Dreyfuss G, Eddy SR, Griffiths-Jones S, Marshall M, et al: A uniform system for microRNA annotation. RNA 2003, 9:277-279. 17. Lagos-Quintana M, Rauhut R, Lendeckel W, Tuschl T: Identification of novel genes coding for small expressed RNAs. Science 2001, 294:853-858. 18. Lagos-Quintana M, Rauhut R, Yalcin A, Meyer J, Lendeckel W, Tuschl T: Identification of tissue-specific microRNAs from mouse. Curr Biol 2002, 12:735-739. 19. Zhao Y, Samal E, Srivastava D: Serum response factor regulates a muscle-specific microRNA that targets Hand2 during cardiogenesis. Nature 2005, 436:214-220. 20. Clarke PG: Developmental cell death: morphological diversity and multiple mechanisms. Anat Embryol (Berl) 1990, 181:195-213. 21. Lockshin RA, Zakeri Z: Apoptosis, autophagy, and more. Int J Biochem Cell Biol 2004, 36:2405-2419. 22. Hotchkiss RS, Strasser A, McDunn JE, Swanson PE: Cell death. N Engl J Med 2009, 361:1570-1583. 23. Vogt C: Untersuchungen über die Entwicklungsgeschichte der Geburtshelferkröte. Solothurn: Jent und Gassman; 1842. 24. Flemming W: Uber die Bildung von Richtungsfiguren in Saugethiereiern beim Untergang Graaf'scher Follikel. Arch Anat EntwGesch 1885:221-244. 25. Kerr JF: Shrinkage necrosis: a distinct mode of cellular death. J Pathol 1971, 105:13-20. 26. Kerr JF, Wyllie AH, Currie AR: Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972, 26:239-257. 27. Kerr JF: A histochemical study of hypertrophy and ischaemic injury of rat liver with special reference to changes in lysosomes. J Pathol Bacteriol 1965, 90:419-435. 28. Majno G, Joris I: Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol 1995, 146:3-15. 29. Van Cruchten S, Van Den Broeck W: Morphological and biochemical aspects of apoptosis, oncosis and necrosis. Anat Histol Embryol 2002, 31:214-223. 30. Enari M, Hug H, Nagata S: Involvement of an ICE-like protease in Fas-mediated apoptosis. Nature 1995, 375:78-81. 31. Kepp O, Galluzzi L, Lipinski M, Yuan J, Kroemer G: Cell death assays for drug discovery. Nat Rev Drug Discov 2011, 10:221-237. 32. Ashford TP, Porter KR: Cytoplasmic components in hepatic cell lysosomes. J Cell Biol 1962, 12:198-202. 33. Klionsky DJ: Autophagy: from phenomenology to molecular understanding in less than a decade. Nat Rev Mol Cell Biol 2007, 8:931-937. 34. Cataldo AM, Hamilton DJ, Barnett JL, Paskevich PA, Nixon RA: Properties of the endosomal-lysosomal system in the human central nervous system: disturbances mark most neurons in populations at risk to degenerate in Alzheimer's disease. J Neurosci 1996, 16:186-199. 35. Ravikumar B, Vacher C, Berger Z, Davies JE, Luo S, Oroz LG, Scaravilli F, Easton DF, Duden R, O'Kane CJ, Rubinsztein DC: Inhibition of mTOR induces autophagy and reduces toxicity of polyglutamine expansions in fly and mouse models of Huntington disease. Nat Genet 2004, 36:585-595. 36. Xilouri M, Stefanis L: Autophagic pathways in Parkinson disease and related disorders. Expert Rev Mol Med 2011, 13:e8. 37. Sasaki S: Autophagy in spinal cord motor neurons in sporadic amyotrophic lateral sclerosis. J Neuropathol Exp Neurol 2011, 70:349-359. 38. von RF: Untersuchungen Ober Rachitis und Osteomalacie. Jena: Verlag Gustav Fischer; 1910. 39. Sanders EJ, Wride MA: Programmed cell death in development. Int Rev Cytol 1995, 163:105-173. 40. Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 2010, 127:2893-2917. 41. Hsu JL, Glaser SL: Epstein-barr virus-associated malignancies: epidemiologic patterns and etiologic implications. Crit Rev Oncol Hematol 2000, 34:27-53. 42. Chien YC, Chen JY, Liu MY, Yang HI, Hsu MM, Chen CJ, Yang CS: Serologic markers of Epstein-Barr virus infection and nasopharyngeal carcinoma in Taiwanese men. N Engl J Med 2001, 345:1877-1882. 43. Raab-Traub N: Epstein-Barr virus in the pathogenesis of NPC. Semin Cancer Biol 2002, 12:431-441. 44. Tsai ST, Jin YT, Mann RB, Ambinder RF: Epstein-Barr virus detection in nasopharyngeal tissues of patients with suspected nasopharyngeal carcinoma. Cancer 1998, 82:1449-1453. 45. Young LS, Rickinson AB: Epstein-Barr virus: 40 years on. Nat Rev Cancer 2004, 4:757-768. 46. zur Hausen H, Schulte-Holthausen H, Klein G, Henle W, Henle G, Clifford P, Santesson L: EBV DNA in biopsies of Burkitt tumours and anaplastic carcinomas of the nasopharynx. Nature 1970, 228:1056-1058. 47. Lin CT, Wong CI, Chan WY, Tzung KW, Ho JK, Hsu MM, Chuang SM: Establishment and characterization of two nasopharyngeal carcinoma cell lines. Lab Invest 1990, 62:713-724. 48. Lin CT, Chan WY, Chen W, Huang HM, Wu HC, Hsu MM, Chuang SM, Wang CC: Characterization of seven newly established nasopharyngeal carcinoma cell lines. Lab Invest 1993, 68:716-727. 49. Haritos AA, Goodall GJ, Horecker BL: Prothymosin alpha: isolation and properties of the major immunoreactive form of thymosin alpha 1 in rat thymus. Proc Natl Acad Sci U S A 1984, 81:1008-1011. 50. Gomez-Marquez J, Rodriguez P: Prothymosin alpha is a chromatin-remodelling protein in mammalian cells. Biochem J 1998, 333 ( Pt 1):1-3. 51. Tsitsiloni OE, Stiakakis J, Koutselinis A, Gogas J, Markopoulos C, Yialouris P, Bekris S, Panoussopoulos D, Kiortsis V, Voelter W, et al.: Expression of alpha-thymosins in human tissues in normal and abnormal growth. Proc Natl Acad Sci U S A 1993, 90:9504-9507. 52. Jiang X, Kim HE, Shu H, Zhao Y, Zhang H, Kofron J, Donnelly J, Burns D, Ng SC, Rosenberg S, Wang X: Distinctive roles of PHAP proteins and prothymosin-alpha in a death regulatory pathway. Science 2003, 299:223-226. 53. Qi X, Wang L, Du F: Novel small molecules relieve prothymosin alpha-mediated inhibition of apoptosome formation by blocking its interaction with Apaf-1. Biochemistry 2010, 49:1923-1930. 54. Shan ZX, Lin QX, Fu YH, Deng CY, Zhou ZL, Zhu JN, Liu XY, Zhang YY, Li Y, Lin SG, Yu XY: Upregulated expression of miR-1/miR-206 in a rat model of myocardial infarction. Biochem Biophys Res Commun 2009, 381:597-601. 55. Xu C, Lu Y, Pan Z, Chu W, Luo X, Lin H, Xiao J, Shan H, Wang Z, Yang B: The muscle-specific microRNAs miR-1 and miR-133 produce opposing effects on apoptosis by targeting HSP60, HSP70 and caspase-9 in cardiomyocytes. J Cell Sci 2007, 120:3045-3052. 56. Tang Y, Zheng J, Sun Y, Wu Z, Liu Z, Huang G: MicroRNA-1 regulates cardiomyocyte apoptosis by targeting Bcl-2. Int Heart J 2009, 50:377-387. 57. Shan ZX, Lin QX, Deng CY, Zhu JN, Mai LP, Liu JL, Fu YH, Liu XY, Li YX, Zhang YY, et al: miR-1/miR-206 regulate Hsp60 expression contributing to glucose-mediated apoptosis in cardiomyocytes. FEBS Lett 2010, 584:3592-3600. 58. Yu XY, Song YH, Geng YJ, Lin QX, Shan ZX, Lin SG, Li Y: Glucose induces apoptosis of cardiomyocytes via microRNA-1 and IGF-1. Biochem Biophys Res Commun 2008, 376:548-552. 59. Datta J, Kutay H, Nasser MW, Nuovo GJ, Wang B, Majumder S, Liu CG, Volinia S, Croce CM, Schmittgen TD, et al: Methylation mediated silencing of MicroRNA-1 gene and its role in hepatocellular carcinogenesis. Cancer Res 2008, 68:5049-5058. 60. Hirai H, Verma M, Watanabe S, Tastad C, Asakura Y, Asakura A: MyoD regulates apoptosis of myoblasts through microRNA-mediated down-regulation of Pax3. J Cell Biol 2010, 191:347-365. 61. Nasser MW, Datta J, Nuovo G, Kutay H, Motiwala T, Majumder S, Wang B, Suster S, Jacob ST, Ghoshal K: Down-regulation of micro-RNA-1 (miR-1) in lung cancer. Suppression of tumorigenic property of lung cancer cells and their sensitization to doxorubicin-induced apoptosis by miR-1. J Biol Chem 2008, 283:33394-33405. 62. Yoshino H, Chiyomaru T, Enokida H, Kawakami K, Tatarano S, Nishiyama K, Nohata N, Seki N, Nakagawa M: The tumour-suppressive function of miR-1 and miR-133a targeting TAGLN2 in bladder cancer. Br J Cancer 2011, 104:808-818. 63. Nohata N, Sone Y, Hanazawa T, Fuse M, Kikkawa N, Yoshino H, Chiyomaru T, Kawakami K, Enokida H, Nakagawa M, et al: miR-1 as a tumor suppressive microRNA targeting TAGLN2 in head and neck squamous cell carcinoma. Oncotarget 2011, 2:29-42. 64. Fujita R, Ueda M, Fujiwara K, Ueda H: Prothymosin-alpha plays a defensive role in retinal ischemia through necrosis and apoptosis inhibition. Cell Death Differ 2009, 16:349-358. 65. Letsas KP, Frangou-Lazaridis M: Surfing on prothymosin alpha proliferation and anti-apoptotic properties. Neoplasma 2006, 53:92-96. 66. Suzuki Y, Imai Y, Nakayama H, Takahashi K, Takio K, Takahashi R: A serine protease, HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death. Mol Cell 2001, 8:613-621. 67. Igaki T, Suzuki Y, Tokushige N, Aonuma H, Takahashi R, Miura M: Evolution of mitochondrial cell death pathway: Proapoptotic role of HtrA2/Omi in Drosophila. Biochem Biophys Res Commun 2007, 356:993-997. 68. Hayashi T, Yoshida S, Yoshinaga A, Ohno R, Ishii N, Yamada T: HtrA2 is up-regulated in the rat testis after experimental cryptorchidism. Int J Urol 2006, 13:157-164. 69. Du C, Fang M, Li Y, Li L, Wang X: Smac, a mitochondrial protein that promotes cytochrome c-dependent caspase activation by eliminating IAP inhibition. Cell 2000, 102:33-42. 70. Verhagen AM, Ekert PG, Pakusch M, Silke J, Connolly LM, Reid GE, Moritz RL, Simpson RJ, Vaux DL: Identification of DIABLO, a mammalian protein that promotes apoptosis by binding to and antagonizing IAP proteins. Cell 2000, 102:43-53. 71. Meulmeester E, Jochemsen AG: p53: a guide to apoptosis. Curr Cancer Drug Targets 2008, 8:87-97. 72. Buckbinder L, Talbott R, Velasco-Miguel S, Takenaka I, Faha B, Seizinger BR, Kley N: Induction of the growth inhibitor IGF-binding protein 3 by p53. Nature 1995, 377:646-649. 73. Miyashita T, Reed JC: Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 1995, 80:293-299. 74. Marchenko ND, Wolff S, Erster S, Becker K, Moll UM: Monoubiquitylation promotes mitochondrial p53 translocation. EMBO J 2007, 26:923-934. 75. Mahmood Z, Shukla Y: Death receptors: targets for cancer therapy. Exp Cell Res 2010, 316:887-899. 76. Mathew SJ, Haubert D, Kronke M, Leptin M: Looking beyond death: a morphogenetic role for the TNF signalling pathway. J Cell Sci 2009, 122:1939-1946. 77. Russo M, Mupo A, Spagnuolo C, Russo GL: Exploring death receptor pathways as selective targets in cancer therapy. Biochem Pharmacol 2010, 80:674-682. 78. Shen HM, Pervaiz S: TNF receptor superfamily-induced cell death: redox-dependent execution. FASEB J 2006, 20:1589-1598. 79. Walczak H, Haas TL: Biochemical analysis of the native TRAIL death-inducing signaling complex. Methods Mol Biol 2008, 414:221-239. 80. Ban L, Zhang J, Wang L, Kuhtreiber W, Burger D, Faustman DL: Selective death of autoreactive T cells in human diabetes by TNF or TNF receptor 2 agonism. Proc Natl Acad Sci U S A 2008, 105:13644-13649. 81. de Oliveira Pinto LM, Garcia S, Lecoeur H, Rapp C, Gougeon ML: Increased sensitivity of T lymphocytes to tumor necrosis factor receptor 1 (TNFR1)- and TNFR2-mediated apoptosis in HIV infection: relation to expression of Bcl-2 and active caspase-8 and caspase-3. Blood 2002, 99:1666-1675. 82. Pan G, Ni J, Wei YF, Yu G, Gentz R, Dixit VM: An antagonist decoy receptor and a death domain-containing receptor for TRAIL. Science 1997, 277:815-818. 83. Pan G, O'Rourke K, Chinnaiyan AM, Gentz R, Ebner R, Ni J, Dixit VM: The receptor for the cytotoxic ligand TRAIL. Science 1997, 276:111-113. 84. Garcia-Ruiz C, Colell A, Mari M, Morales A, Calvo M, Enrich C, Fernandez-Checa JC: Defective TNF-alpha-mediated hepatocellular apoptosis and liver damage in acidic sphingomyelinase knockout mice. J Clin Invest 2003, 111:197-208. 85. Korner H, Sedgwick JD: Tumour necrosis factor and lymphotoxin: molecular aspects and role in tissue-specific autoimmunity. Immunol Cell Biol 1996, 74:465-472. 86. Lu G, Janjic BM, Janjic J, Whiteside TL, Storkus WJ, Vujanovic NL: Innate direct anticancer effector function of human immature dendritic cells. II. Role of TNF, lymphotoxin-alpha(1)beta(2), Fas ligand, and TNF-related apoptosis-inducing ligand. J Immunol 2002, 168:1831-1839. 87. Obeid LM, Linardic CM, Karolak LA, Hannun YA: Programmed cell death induced by ceramide. Science 1993, 259:1769-1771. 88. Oliveira RB, Sampaio EP, Aarestrup F, Teles RM, Silva TP, Oliveira AL, Antas PR, Sarno EN: Cytokines and Mycobacterium leprae induce apoptosis in human Schwann cells. J Neuropathol Exp Neurol 2005, 64:882-890. 89. Huang H, Joazeiro CA, Bonfoco E, Kamada S, Leverson JD, Hunter T: The inhibitor of apoptosis, cIAP2, functions as a ubiquitin-protein ligase and promotes in vitro monoubiquitination of caspases 3 and 7. J Biol Chem 2000, 275:26661-26664. 90. Roy N, Deveraux QL, Takahashi R, Salvesen GS, Reed JC: The c-IAP-1 and c-IAP-2 proteins are direct inhibitors of specific caspases. EMBO J 1997, 16:6914-6925. 91. Deveraux QL, Roy N, Stennicke HR, Van Arsdale T, Zhou Q, Srinivasula SM, Alnemri ES, Salvesen GS, Reed JC: IAPs block apoptotic events induced by caspase-8 and cytochrome c by direct inhibition of distinct caspases. EMBO J 1998, 17:2215-2223. 92. Owens TW, Foster FM, Valentijn A, Gilmore AP, Streuli CH: Role for X-linked Inhibitor of apoptosis protein upstream of mitochondrial permeabilization. J Biol Chem 2010, 285:1081-1088. 93. Dallman C, Johnson PW, Packham G: Differential regulation of cell survival by CD40. Apoptosis 2003, 8:45-53. 94. Chandra D, Tang DG: Mitochondrially localized active caspase-9 and caspase-3 result mostly from translocation from the cytosol and partly from caspase-mediated activation in the organelle. Lack of evidence for Apaf-1-mediated procaspase-9 activation in the mitochondria. J Biol Chem 2003, 278:17408-17420. 95. Kim SC, Stice JP, Chen L, Jung JS, Gupta S, Wang Y, Baumgarten G, Trial J, Knowlton AA: Extracellular heat shock protein 60, cardiac myocytes, and apoptosis. Circ Res 2009, 105:1186-1195. 96. Lin L, Kim SC, Wang Y, Gupta S, Davis B, Simon SI, Torre-Amione G, Knowlton AA: HSP60 in heart failure: abnormal distribution and role in cardiac myocyte apoptosis. Am J Physiol Heart Circ Physiol 2007, 293:H2238-2247. 97. Kirchhoff SR, Gupta S, Knowlton AA: Cytosolic heat shock protein 60, apoptosis, and myocardial injury. Circulation 2002, 105:2899-2904. 98. Saleh A, Srinivasula SM, Balkir L, Robbins PD, Alnemri ES: Negative regulation of the Apaf-1 apoptosome by Hsp70. Nat Cell Biol 2000, 2:476-483. 99. Ravagnan L, Gurbuxani S, Susin SA, Maisse C, Daugas E, Zamzami N, Mak T, Jaattela M, Penninger JM, Garrido C, Kroemer G: Heat-shock protein 70 antagonizes apoptosis-inducing factor. Nat Cell Biol 2001, 3:839-843. 100. Park HS, Cho SG, Kim CK, Hwang HS, Noh KT, Kim MS, Huh SH, Kim MJ, Ryoo K, Kim EK, et al: Heat shock protein hsp72 is a negative regulator of apoptosis signal-regulating kinase 1. Mol Cell Biol 2002, 22:7721-7730. 101. Stankiewicz AR, Lachapelle G, Foo CP, Radicioni SM, Mosser DD: Hsp70 inhibits heat-induced apoptosis upstream of mitochondria by preventing Bax translocation. J Biol Chem 2005, 280:38729-38739. 102. Ambrosini G, Adida C, Altieri DC: A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med 1997, 3:917-921. 103. Kim YS, Koh JM, Lee YS, Kim BJ, Lee SH, Lee KU, Kim GS: Increased circulating heat shock protein 60 induced by menopause, stimulates apoptosis of osteoblast-lineage cells via up-regulation of toll-like receptors. Bone 2009, 45:68-76. 104. Oh HK, Tan AL, Das K, Ooi CH, Deng NT, Tan IB, Beillard E, Lee J, Ramnarayanan K, Rha SY, et al: Genomic loss of miR-486 regulates tumor progression and the OLFM4 antiapoptotic factor in gastric cancer. Clin Cancer Res 2011, 17:2657-2667. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66666 | - |
dc.description.abstract | 微核醣核酸 (microRNA) 是細胞內自行轉錄產生長度約 21 個核苷酸的非編碼核醣核酸 (non-coding RNA)。 microRNA 的功能是負向調控其它基因的表現及在各種細胞功能中扮演重要的角色。微核醣核酸-1 (miR-1) 常被使用在 microRNA 實驗中當正對照組。先前在我們其它 microRNA 的實驗中,當 miR-1 加到鼻咽癌細胞株 NPC-TW01 中當作正對照組,在曠時攝影顯微鏡觀察下出現很典型的細胞凋亡 (apoptosis) 現象。進一步用Annexin V、TUNEL 染色及caspase assay證實 miR-1會引發NPC-TW01 鼻咽癌細胞株的細胞凋亡。 測試其它細胞株,發現 miR-1也會讓 HeLa、 Cal-27、 KYSE-30及NPC-TW06等原本細胞內就低度表現miR-1的細胞株產生細胞凋亡, 但 miR-1不會讓 HepG2、 SW620、 HEK-293T、 SAS 等原本細胞內就高度表現miR-1的細胞株產生細胞凋亡。 為了解 miR-1誘導細胞凋亡的分子機制,我們找出 miR-1直接調控並且參與細胞凋亡作用的基因。方法是首先將miR-1送入NPC-TW01 與 HeLa 細胞中,並用 cDNA 表現微陣列(cDNA expression microarray) 篩選會被 miR-1抑制表現的基因,這些基因再到 microRNA 資料庫去篩選可能是 miR-1直接作用的標的基因 (target gene),最後再進到細胞訊息傳遞資料庫(Ingenuity pathway database) 篩選與細胞凋亡有關的基因。總共有C5、CARD8、FAIM、GRIN2A、TP63及PTMA等6個基因符合條件。再利用Q-RT-PCR及冷光報導基因載體 (luciferase reporter vector) 方法證實miR-1會直接調控PTMA基因。因 miR-1會抑制PTMA的表現且會誘導細胞凋亡,所以我們使用PTMA siRNA代替 miR-1來抑制 PTMA的表現,並觀察PTMA siRNA 抑制PTMA表現後是否也會誘導細胞凋亡。結果單獨抑制PTMA的表現並不會誘導細胞凋亡,但會加速細胞凋亡誘導劑 (apoptotic inducer) 處理過的細胞進行細胞凋亡作用。此外我們使用細胞凋亡抗體陣列 (apoptosis antibody array) 發現miR-1會誘導一些促細胞凋亡蛋白 (pro-apoptotic protein) 的表現,及抑制抗細胞凋亡蛋白 (anti-apoptotic protein) 的表現。 總結本研究證明 miR-1會誘導鼻咽癌及其它癌細胞株進行細胞凋亡。這是一個新的microRNA誘導細胞凋亡的模式。它的機制是miR-1透過直接調控PTMA mRNA及間接調控其它細胞凋亡蛋白方式來進行。 PTMA siRNA會加速被細胞凋亡誘導劑處理過的細胞進行細胞凋亡,因有些細胞凋亡誘導劑 (例如Actinomycin D, camptothecin 及 etoposide) 在臨床上也是癌症化療藥物,所以PTMA siRNA在癌症治療上有當成化療輔助劑的潛在應用價值。 此外我們研究NPC-TW01與NPC-TW01N1細胞的腫瘤生成能力差異時,找到miR-486在兩個細胞間表現有差別。將miR-486送入原本細胞內很少表現miR-486的NPC-TW01N1細胞內,結果會降低NPC-TW01N1細胞的增殖速率及引起細胞凋亡。 | zh_TW |
dc.description.abstract | MicroRNAs are endogenous non-coding single-strand RNAs, approximately 21 nucleotides in size. MicroRNAs function as negative regulators of gene expression and play important roles in biological processes. MicroRNA-1 (miR-1) has been used as a positive control in some microRNA experiments. Previously in our other microRNA experiment, when miR-1 was transfected into nasopharyngeal carcinoma (NPC-TW01) cells as a positive control, to our surprise, a typical apoptotic process was observed in NPC cells by time-lapse microscopy. Further observations using Annexin V staining, TUNEL staining, caspase assay all confirmed that miR-1 could induce NPC cell apoptosis. Transfection of miR-1 into other cancer cell lines, such as the HeLa, Cal-27, KYSE30, and NPC-TW06 which express low levels of endogenous miR-1, also induced apoptosis, but certain another cancer cell lines, such as SW620, HepG2, SAS, and PC-13 which express high levels of endogenous miR-1, did not show apoptosis. In order to clear the molecular mechanism of miR-1 inducing apoptosis, and identify the miR-1 directly regulated genes that involve apoptosis, cDNA expression microarray analysis of miR-1 transfected NPC-TW01 and HeLa cells were performed. All the down regulated genes were analyzed through microRNA database and Ingenuity pathway database. Six candidate miR-1 regulated genes, C5, CARD8, FAIM, GRIN2A, TP63, and PTMA were found. Using Q-RT-PCR analysis and luciferase reporter assay demonstrate that miR-1 could directly target PTMA mRNA. MiR-1 can induce cell apoptosis and directly down regulate PTMA mRNA so we used PTMA siRNA in place of miR-1 to down regulate PTMA mRNA and observed the apoptotic effect. We found that down regulated PTMA alone could not induce apoptosis, but PTMA siRNA could accelerate the apoptotic process when cells were treated with apoptotic inducers. Furthermore, using apoptosis antibody array, we found that miR-1 could up regulated some pro-apoptotic protein expressions and down-regulated anti-apoptotic protein expressions. We conclude that miR-1 could induce apoptosis in NPC and some other cancer cell lines. This is a novel model of microRNA-induced cell apoptosis. The mechanism is through miR-1 direct targeting of the apoptotic inhibitor PTMA mRNA and through indirect regulation of other apoptotic protein expressions. The PTMA siRNA can accelerate the apoptotic process when cells are treated with apoptotic inducers. The apoptotic inducers: actinomycin D, camptothecin and etoposide are also as chemotherapy drugs in clinical cancer therapy so PTMA siRNA may have potential applications as an adjuvant in cancer chemotherapy. In addition, we investigate the tumorigenesis variation between NPC-TW01 and NPC-TW01N1 and found miR-486 is alteration expression. Transfection of miR-486 into NPC-TW01N1 which expression low levels of endogenous miR-486 can decrease cell proliferation rate and induce cell apoptosis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T00:49:55Z (GMT). No. of bitstreams: 1 ntu-100-D90444003-1.pdf: 4567544 bytes, checksum: 697f16ec72682a65aeea24ee4bc4a69d (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | Abstract in Chinese………………………………………………………………………I
Abstract in English………………………………………………………………..……III Abbreviation……………………………………………………………………………VI Contents……………………….………………………………………………………VII Chapter 1. Introduction…………………………………………………………….……1 Chapter 2. Materials and Methods………………….……………………….…………10 Cell lines…………………………………………………………………….……10 Transfection of microRNA and siRNA……………………………..……………10 Apoptosis assay………………………………………………………………...…11 Microarray analysis………………………………………………………….……12 Quantitative RT–PCR…………….................................................................……13 Luciferase reporter assay……………............................................................……13 Animal experiments………………………………………………………………14 Chapter 3. results………………………………………………………………………15 Part I: miR-1 induces apoptosis………………………………………………..………15 MiR-1 induces NPC cells apoptosis………………………………………………15 MiR-1 directly targets PTMA………………………………………………….…17 PTMA assisting cells proceed to apoptosis…………………………….…………18 MiR-1 influences some apoptotic protein expressions……………………………20 Part II: miR-486 involves tumorigenesis by inhibiting cell proliferation and inducing cell apoptosis………………………………………………………………………...…21 The tumorigenesis capability of NPC-TW01N1 is more than NPC-TW01………21 Comparing the phenotype between NPC-TW01 and NPC-TW01N1 cells……….21 Comparing the genotype between NPC-TW01 and NPC-TW01N1 cells………...23 MiR-486 decreases NPC-TW01N1 proliferation rate and induces apoptosis……23 Chapter 4. Discussion and Conclusion…………………………………………………24 Chapter 5. References………………………………………………………………..…32 Figures…………………………………………………………………………………41 Figure 1 Transfection of miR-1 into NPC-TW01 cells causes cell death……...…42 Figure 2 Transfection of miR-1 into NPC-TW01 cells appear apoptotic bodies…43 Figure 3 Transfection of miR-1 into NPC-TW01 cell appear nuclear fragmentation ……………………………………………………………44 Figure 4 Annexin V analyses miR-1 inducing apoptotic cells…………………....45 Figure 5 TUNEL assay of the fragmented DNA from miR-1 inducing apoptotic cells………………………………………………………………………47 Figure 6 Transfection of miR-1 into Cal-27 and SW620 cells then observe their morphology………………………………………………………………48 Figure 7 Luciferase assay demonstrates that miR-1 directly targets PTMA 3’ UTR………………………………………………………………..……49 Figure 8 PTMA siRNA and miR-1 accelerate the apoptotic process in cells that were treated with actinomycin D…………….………………….………51 Figure 9 Apoptotic protein profiles of NPC-TW01 and HeLa cells transfected with miR-1………………………………………………………………….…53 Figure 10 Schematic representation of the relationship among miR-1, PTMA during apoptosis…………………………………………………………55 Figure 11 Schematic representation of the generation of NPC-TW01 and NPC-TW01N1 cell lines…………………………………………………56 Figure 12 The comparison of tumorigenesis capability between NPC-TW01 and NPC-TW01N1………………………………………………………...…57 Figure 13 The comparison of DNA profile (STR) between NPC-TW01 and NPC-TW01N1………………………………………………………..…59 Figure 14 The comparison of morphology between NPC-TW01 and NPC-TW01N1 cells…………………………………………………...…60 Figure 15 The comparison of proliferation capability between NPC-TW01 and NPC-TW01N1 cells…………………………………………………...…61 Figure 16 The comparison of cell death between NPC-TW01 and NPC-TW01N1 cells………………………………………………………………………63 Figure 17 The comparison of angiogenesis capability between NPC-TW01 and NPC-TW01N1………………………………………………………..…64 Figure 18 The comparison of invasion capability between NPC-TW01 and NPC-TW01N1 cells………………………………………………….…65 Figure 19 The comparison of migration capability between NPC-TW01 and NPC-TW01N1 cells………………………………………………….…66 Figure 20 Transfection of miR-486 decreases NPC-TW01N1 cells proliferation rate and induces apoptosis………………………………………………67 Figure 21 Transfection of miR-486 increases NPC-TW01N1 cells death….……69 Table……………………………………………………………………………..……70 Table 1 Candidate targeting genes of miR-1 that involve apoptosis….…………70 Table 2 Q-RT-PCR analysis confirming miR-1 candidate targeting genes.…..…71 Table 3 Primers used for Q-RT-PCR……………………………………….……72 Table 4 Oligos used for pmirGLO luciferase reporter vector construction………73 Table 5 Comparison the endogenous microRNA express levels between NPC-TW01 and NPC-TW01N1…………………………………………74 Appendix published articles…………………………………………………….……...75 | |
dc.language.iso | en | |
dc.title | 微核醣核酸誘導鼻咽癌細胞株細胞凋亡之機制 | zh_TW |
dc.title | The mechanism of microRNA inducing apoptosis in nasopharyngeal carcinoma cell lines | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-1 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 蘇燦隆,吳漢忠,林中梧,李明學,王萬波 | |
dc.subject.keyword | 鼻咽癌,微核醣核酸,前胸腺素α,細胞凋亡,微核醣核酸-1,微核醣核酸-486, | zh_TW |
dc.subject.keyword | Nasopharyngeal carcinoma (NPC),microRNA,PTMA (prothymosin alpha,ProTalpha),apoptosis,miR-1,miR-486, | en |
dc.relation.page | 76 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-11-29 | |
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-100-1.pdf 目前未授權公開取用 | 4.46 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。