請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65356
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 周文堅 | |
dc.contributor.author | Shih-Chiang Lin | en |
dc.contributor.author | 林世強 | zh_TW |
dc.date.accessioned | 2021-06-16T23:38:19Z | - |
dc.date.available | 2012-09-19 | |
dc.date.copyright | 2012-09-19 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-07-25 | |
dc.identifier.citation | 1. Jones, P.A. and S.B. Baylin, The epigenomics of cancer. Cell, 2007. 128(4): p. 683-92.
2. Sharma, S., T.K. Kelly, and P.A. Jones, Epigenetics in cancer. Carcinogenesis, 2010. 31(1): p. 27-36. 3. Vogelstein, B. and K.W. Kinzler, The multistep nature of cancer. Trends Genet, 1993. 9(4): p. 138-41. 4. Visani, G., et al., The prognostic value of cytogenetics is reinforced by the kind of induction/consolidation therapy in influencing the outcome of acute myeloid leukemia--analysis of 848 patients. Leukemia, 2001. 15(6): p. 903-9. 5. Esteller, M., Epigenetic gene silencing in cancer: the DNA hypermethylome. Hum Mol Genet, 2007. 16 Spec No 1: p. R50-9. 6. Jenuwein, T. and C.D. Allis, Translating the histone code. Science, 2001. 293(5532): p. 1074-80. 7. Ellis, L., P.W. Atadja, and R.W. Johnstone, Epigenetics in cancer: targeting chromatin modifications. Mol Cancer Ther, 2009. 8(6): p. 1409-20. 8. Wang, G.G., C.D. Allis, and P. Chi, Chromatin remodeling and cancer, Part I: Covalent histone modifications. Trends Mol Med, 2007. 13(9): p. 363-72. 9. Bannister, A.J. and T. Kouzarides, Regulation of chromatin by histone modifications. Cell Res, 2011. 21(3): p. 381-95. 10. Guil, S. and M. Esteller, DNA methylomes, histone codes and miRNAs: tying it all together. Int J Biochem Cell Biol, 2009. 41(1): p. 87-95. 11. Cedar, H. and Y. Bergman, Linking DNA methylation and histone modification: patterns and paradigms. Nat Rev Genet, 2009. 10(5): p. 295-304. 12. Hashimshony, T., et al., The role of DNA methylation in setting up chromatin structure during development. Nat Genet, 2003. 34(2): p. 187-92. 13. Lindroth, A.M., et al., Antagonism between DNA and H3K27 methylation at the imprinted Rasgrf1 locus. PLoS Genet, 2008. 4(8): p. e1000145. 14. Bird, A.P., CpG-rich islands and the function of DNA methylation. Nature, 1986. 321(6067): p. 209-13. 15. Bird, A., DNA methylation patterns and epigenetic memory. Genes Dev, 2002. 16(1): p. 6-21. 16. Barciszewska, A.M., et al., Analysis of 5-methylcytosine in DNA of breast and colon cancer tissues. IUBMB Life, 2007. 59(12): p. 765-70. 17. Jones, P.A. and S.B. Baylin, The fundamental role of epigenetic events in cancer. Nat Rev Genet, 2002. 3(6): p. 415-28. 18. Weber, M., et al., Chromosome-wide and promoter-specific analyses identify sites of differential DNA methylation in normal and transformed human cells. Nat Genet, 2005. 37(8): p. 853-62. 19. Yoo, C.B. and P.A. Jones, Epigenetic therapy of cancer: past, present and future. Nat Rev Drug Discov, 2006. 5(1): p. 37-50. 20. Kriaucionis, S. and N. Heintz, The nuclear DNA base 5-hydroxymethylcytosine is present in Purkinje neurons and the brain. Science, 2009. 324(5929): p. 929-30. 21. Song, C.X., et al., Selective chemical labeling reveals the genome-wide distribution of 5-hydroxymethylcytosine. Nat Biotechnol, 2011. 29(1): p. 68-72. 22. Valinluck, V. and L.C. Sowers, Endogenous cytosine damage products alter the site selectivity of human DNA maintenance methyltransferase DNMT1. Cancer Res, 2007. 67(3): p. 946-50. 23. Pastor, W.A., et al., Genome-wide mapping of 5-hydroxymethylcytosine in embryonic stem cells. Nature, 2011. 473(7347): p. 394-7. 24. Tahiliani, M., et al., Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1. Science, 2009. 324(5929): p. 930-5. 25. Ko, M., et al., Impaired hydroxylation of 5-methylcytosine in myeloid cancers with mutant TET2. Nature, 2010. 468(7325): p. 839-43. 26. Chou, W.C., et al., TET2 mutation is an unfavorable prognostic factor in acute myeloid leukemia patients with intermediate-risk cytogenetics. Blood, 2011. 118(14): p. 3803-10. 27. Ito, S., et al., Tet proteins can convert 5-methylcytosine to 5-formylcytosine and 5-carboxylcytosine. Science, 2011. 333(6047): p. 1300-3. 28. Halkidou, K., et al., Upregulation and nuclear recruitment of HDAC1 in hormone refractory prostate cancer. Prostate, 2004. 59(2): p. 177-89. 29. Peters, A.H., et al., Loss of the Suv39h histone methyltransferases impairs mammalian heterochromatin and genome stability. Cell, 2001. 107(3): p. 323-37. 30. Fraga, M.F., et al., Loss of acetylation at Lys16 and trimethylation at Lys20 of histone H4 is a common hallmark of human cancer. Nat Genet, 2005. 37(4): p. 391-400. 31. Muller, J., et al., Histone methyltransferase activity of a Drosophila Polycomb group repressor complex. Cell, 2002. 111(2): p. 197-208. 32. Czermin, B., et al., Drosophila enhancer of Zeste/ESC complexes have a histone H3 methyltransferase activity that marks chromosomal Polycomb sites. Cell, 2002. 111(2): p. 185-96. 33. Cao, R., et al., Role of histone H3 lysine 27 methylation in Polycomb-group silencing. Science, 2002. 298(5595): p. 1039-43. 34. Kuzmichev, A., et al., Histone methyltransferase activity associated with a human multiprotein complex containing the Enhancer of Zeste protein. Genes Dev, 2002. 16(22): p. 2893-905. 35. Simon, J.A. and C.A. Lange, Roles of the EZH2 histone methyltransferase in cancer epigenetics. Mutat Res, 2008. 647(1-2): p. 21-9. 36. Ringrose, L. and R. Paro, Epigenetic regulation of cellular memory by the Polycomb and Trithorax group proteins. Annu Rev Genet, 2004. 38: p. 413-43. 37. Su, I.H., et al., Ezh2 controls B cell development through histone H3 methylation and Igh rearrangement. Nat Immunol, 2003. 4(2): p. 124-31. 38. Heard, E., Delving into the diversity of facultative heterochromatin: the epigenetics of the inactive X chromosome. Curr Opin Genet Dev, 2005. 15(5): p. 482-9. 39. Varambally, S., et al., The polycomb group protein EZH2 is involved in progression of prostate cancer. Nature, 2002. 419(6907): p. 624-9. 40. van Galen, J.C., et al., Distinct expression patterns of polycomb oncoproteins and their binding partners during the germinal center reaction. Eur J Immunol, 2004. 34(7): p. 1870-81. 41. Velichutina, I., et al., EZH2-mediated epigenetic silencing in germinal center B cells contributes to proliferation and lymphomagenesis. Blood, 2010. 116(24): p. 5247-55. 42. Kleer, C.G., et al., EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cells. Proc Natl Acad Sci U S A, 2003. 100(20): p. 11606-11. 43. Fluge, O., et al., Expression of EZH2 and Ki-67 in colorectal cancer and associations with treatment response and prognosis. Br J Cancer, 2009. 101(8): p. 1282-9. 44. Wei, Y., et al., Loss of trimethylation at lysine 27 of histone H3 is a predictor of poor outcome in breast, ovarian, and pancreatic cancers. Mol Carcinog, 2008. 47(9): p. 701-6. 45. He, L.R., et al., Prognostic impact of H3K27me3 expression on locoregional progression after chemoradiotherapy in esophageal squamous cell carcinoma. BMC Cancer, 2009. 9: p. 461. 46. Morin, R.D., et al., Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin. Nat Genet, 2010. 42(2): p. 181-5. 47. Yap, D.B., et al., Somatic mutations at EZH2 Y641 act dominantly through a mechanism of selectively altered PRC2 catalytic activity, to increase H3K27 trimethylation. Blood, 2011. 117(8): p. 2451-9. 48. Park, S.W., et al., Mutational analysis of EZH2 codon 641 in non-Hodgkin lymphomas and leukemias. Leuk Res, 2011. 35(1): p. e6-7. 49. Huang, S.F., et al., High frequency of epidermal growth factor receptor mutations with complex patterns in non-small cell lung cancers related to gefitinib responsiveness in Taiwan. Clin Cancer Res, 2004. 10(24): p. 8195-203. 50. Sander, S., et al., MYC stimulates EZH2 expression by repression of its negative regulator miR-26a. Blood, 2008. 112(10): p. 4202-12. 51. Visser, H.P., et al., The Polycomb group protein EZH2 is upregulated in proliferating, cultured human mantle cell lymphoma. Br J Haematol, 2001. 112(4): p. 950-8. 52. Sasaki, D., et al., Overexpression of Enhancer of zeste homolog 2 with trimethylation of lysine 27 on histone H3 in adult T-cell leukemia/lymphoma as a target for epigenetic therapy. Haematologica, 2011. 96(5): p. 712-9. 53. A predictive model for aggressive non-Hodgkin's lymphoma. The International Non-Hodgkin's Lymphoma Prognostic Factors Project. N Engl J Med, 1993. 329(14): p. 987-94. 54. Alizadeh, A.A., et al., Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling. Nature, 2000. 403(6769): p. 503-11. 55. Hans, C.P., et al., Confirmation of the molecular classification of diffuse large B-cell lymphoma by immunohistochemistry using a tissue microarray. Blood, 2004. 103(1): p. 275-82. 56. Winter, J.N., et al., Expression of p21 protein predicts clinical outcome in DLBCL patients older than 60 years treated with R-CHOP but not CHOP: a prospective ECOG and Southwest Oncology Group correlative study on E4494. Clin Cancer Res, 2010. 16(8): p. 2435-42. 57. Mounier, N., et al., Rituximab plus CHOP (R-CHOP) overcomes bcl-2--associated resistance to chemotherapy in elderly patients with diffuse large B-cell lymphoma (DLBCL). Blood, 2003. 101(11): p. 4279-84. 58. Winter, J.N., et al., Prognostic significance of Bcl-6 protein expression in DLBCL treated with CHOP or R-CHOP: a prospective correlative study. Blood, 2006. 107(11): p. 4207-13. 59. Wolk, M. and J.E. Martin, Titrimetric immunohistochemical evaluation of DNA hypomethylation in uterine tumours. J Clin Pathol, 2009. 62(11): p. 1039-42. 60. Ernst, T., et al., Inactivating mutations of the histone methyltransferase gene EZH2 in myeloid disorders. Nat Genet, 2010. 42(8): p. 722-6. 61. Sehn, L.H., et al., Introduction of combined CHOP plus rituximab therapy dramatically improved outcome of diffuse large B-cell lymphoma in British Columbia. J Clin Oncol, 2005. 23(22): p. 5027-33. 62. Philip, T., et al., Autologous bone marrow transplantation as compared with salvage chemotherapy in relapses of chemotherapy-sensitive non-Hodgkin's lymphoma. N Engl J Med, 1995. 333(23): p. 1540-5. 63. Tan, L. and Y.G. Shi, Tet family proteins and 5-hydroxymethylcytosine in development and disease. Development, 2012. 139(11): p. 1895-902. 64. Haffner, M.C., et al., Global 5-hydroxymethylcytosine content is significantly reduced in tissue stem/progenitor cell compartments and in human cancers. Oncotarget, 2011. 2(8): p. 627-37. 65. Kudo, Y., et al., Loss of 5-hydroxymethylcytosine is accompanied with malignant cellular transformation. Cancer Sci, 2012. 103(4): p. 670-6. 66. Yang, H., et al., Tumor development is associated with decrease of TET gene expression and 5-methylcytosine hydroxylation. Oncogene, 2012. 67. Bracken, A.P., et al., EZH2 is downstream of the pRB-E2F pathway, essential for proliferation and amplified in cancer. EMBO J, 2003. 22(20): p. 5323-35. 68. Johnson, N.A., et al., Lymphomas with concurrent BCL2 and MYC translocations: the critical factors associated with survival. Blood, 2009. 114(11): p. 2273-9. 69. Shen, X., et al., EZH1 mediates methylation on histone H3 lysine 27 and complements EZH2 in maintaining stem cell identity and executing pluripotency. Mol Cell, 2008. 32(4): p. 491-502. 70. Dudziec, E., et al., Integrated epigenome profiling of repressive histone modifications, DNA methylation and gene expression in normal and malignant urothelial cells. PLoS One, 2012. 7(3): p. e32750. 71. Booth, M.J., et al., Quantitative sequencing of 5-methylcytosine and 5-hydroxymethylcytosine at single-base resolution. Science, 2012. 336(6083): p. 934-7. 72. Ryan, R.J., et al., EZH2 codon 641 mutations are common in BCL2-rearranged germinal center B cell lymphomas. PLoS One, 2011. 6(12): p. e28585. 73. Zhou, J., et al., The histone methyltransferase inhibitor, DZNep, up-regulates TXNIP, increases ROS production, and targets leukemia cells in AML. Blood, 2011. 118(10): p. 2830-9. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65356 | - |
dc.description.abstract | 傳統上,淋巴癌病人對化學治療反應效果的預測只能依靠International Prognostic Index (IPI score,國際預後索引),目前並沒有更加實用且準確的分子生物層次方面的預測因子可以使用。我們希望藉由表觀基因學 (epigenetic) 在淋巴癌上的研究來找到新的預後因子。
EZH2蛋白的過度表現與Histone 3-lysine 27三個甲基化 (H3K27me3)的減少,在乳癌、攝護腺癌等很多癌症上是一個預後變差的因子。我們的研究主要是探討EZH2蛋白,它具有甲基轉化酶的功能而且會去增加H3K27的甲基化,並藉由這個位置上三個甲基化的程度來調控基因的功能。在癌細胞上,抑癌基因的過度甲基化會強化其DNA與組蛋白的纏繞結合,進而減低抑癌基因之表達,最後導致癌症惡性度變高。 在Cytosine的5-position加上一個甲基就是 5-methylcytosine (5-mC),它是表觀基因學上一個重要的標記。我們可以偵測 5-mC的量來代表整個DNA甲基化的程度。DNA甲基化的改變會導致人類自體免疫的疾病、神經學的病變與癌症,而癌細胞一般會呈現DNA整體去甲基化與局部基因的過度甲基化。而5-mC的hydroxylated型態則是5-hydroxymethylcytosine (5-hmC),他在動物的中樞神經系統含量較高,此標記可能代表DNA去甲基化的程度。在Tahiliani (2010)的研究中,5-mC可以被Ten-Eleven-Translocation (TET) protein轉換成5-hmC,會去改變甲基化的程度;在TET1減少時,會增加基因的甲基化。在急性骨髓性血癌中發現TET2的突變而減少5-hmC的量,此結果在臨床上則會使急性骨髓性血癌的預後變差,而在瀰漫性大型B細胞淋巴癌(DLBCL)病人的檢體中則無相關的研究。除此之外,在老鼠的胚胎幹細胞中發現,5-hmC會特別豐富的存在於基因起始子含有H3K27me3與H3K4me3的地方,5-hmC也被推測可能有暫停轉錄的作用。 在西方國家的研究中發現,21.7% 的germinal center B cell (GCB) 類型的DLBCL中有EZH2 Tyr 641的體細胞突變,在體外試驗中發現此突變蛋白會選擇性的使甲基化的功能增加。然而,在DLBCL族群中,EZH2蛋白的過度表現、增加H3K27三個甲基化與整個DNA甲基化的程度並沒有被完整的研究。所以我的研究是採用回溯性的研究方法,先針對淋巴癌病患的病理檢體來進行研究分析,再與病人之前的治療效果與臨床表現作相關性的探討。本研究主要是希望找出分子生物層次方面的預後因子,進而改善未來淋巴癌病人治療的效果。再來,瞭解5-mC/ 5-hmC/ EZH2/ H3k27me3在淋巴癌檢體中的相關性。最後,分析EZH2 Tyr 641在亞洲的淋巴癌病患中的突變情形。 本實驗是將之前診斷為DLBCL的107位亞東紀念醫院與83位奇美醫院病患之病理檢體做兩個部份的處理:第一部份將病理檢體做特殊染色Immunohistochemistry (IHC) stain,包含anti-H3K27me3、anti-EZH2、anti-5-mC 與anti-5-hmC來定量病人檢體Histone甲基化,EZH2的表現與整體DNA甲基化的狀況; 第二部份是將此病理檢體之DNA萃取出來,來檢驗EZH2上Tyr641的突變狀態。之後做病歷資料的回顧,分析病理EZH2/ Histone methylation/ DNA methylation/ DNA hydroxymethylation/ Tyr641 mutation的表現與病人癌症的臨床狀況,進而瞭解這些基因與蛋白是否可用為預測淋巴癌病人預後的因子,最後用來改善未來淋巴癌病人治療的效果。 目前從西元2002年至2010年底的病理報告中初步篩選出190位確定診斷DLBCL的病患,只用接受標準化療(CHOP或R-CHOP)的138位病人進行生存分析。其中90位是男性,48位是女性,年紀最輕是13歲,年紀最大是86歲,平均年齡為57.2歲。癌症分期中,第一與第二期的病患最多。IPI score則是以0~2分的最多。治療的處方以R-CHOP這種標準處方的病人最多。經第一線化學治療後,達到完全緩解(complete remission, CR)的病人佔80%。平均追蹤時間是42個月,而病人的存活時間,最長3475天,最短39天(從診斷到死亡)。 初步收集的48位病人的檢體並無EZH2 基因之hot spot mutation,與文獻中的狀況不相同。在病患的臨床資料與四種IHC染色的分析中觀察到,H3K27me3的強弱表現與性別的分布有顯著的差異(P=0.044),而5-mC的強弱也與治療的反應有相關性(P=0.006),其他因子皆與四種IHC染色無相關性。而四種IHC染色中,H3K27me3與5-mC以及5-mC與5-hmC彼此間有正相關性。 在單變項分析下,stage、IPI score與化學治療的反應對overall survival (OS)有統計學上的意義,而四種抗體並無對OS有統計學上的意義,只有5-hmC有trend(P=0.061)。在多變項分析下,四種抗體個別的強弱仍沒有對OS有統計學上的意義。接著用H3K27me3搭配5-hmC來做分組,其中high H3K27me3/ low 5-hmC的病人存活率較高而high H3K27me3/ high 5-hmC的病人存活率較低(P=0.0497)。在多變項分析後,H3K27me3/5-hmC的分組就沒有統計學上的意義。 在subgroup 分析中進一步發現,在經標準化學治療後CR的110位病人中,H3K27me3/5-hmC的分組就對OS有統計學上的意義(P=0.013)。經多變項分析後,這種分類仍然有統計學上的意義(P=0.017),其中high H3K27me3/ low 5-hmC的病人存活率較高。 結論:我們的研究中指出在DLBCL中H3K27me3/5-hmC的分類可能與預後有關,特別是在CR的族群中,這個high H3K27me3/ low 5-hmC表現的病人存活率較高。四種IHC染色中,H3K27me3與5-mC以及5-mC與5-hmC彼此間有正相關性。而EZH2 Tyr641突變的比例在我們台灣的這個族群的確偏低。 | zh_TW |
dc.description.abstract | Purpose
International Prognostic Index (IPI) score is used to predict the prognosis in diffuse large B cell lymphoma (DLBCL) for around 20 years. However, few molecular prognostic factors were proposed. Overexpression of Enhancer of zeste homolog 2 (EZH2) and decreased histone 3-lysine 27 tri-methylation (H3K27me3) are associated with poor prognosis in many cancers. Promoter CpG island hypermethylation of tumor-suppressor genes is a common hallmark of all human cancers. 5-methylcytosine (5-mC) levels in cancer cells can reflect the DNA hypermethylation status and measurement of 5-hydroxymethylcytosine (5-hmC) levels may estimate the DNA demethylation status in cancer cells. A unique EZH2 Tyr 641 somatic mutation was detected in DLBCL and follicular lymphoma. However, the clinical implications of DNA methylation status, DNA hydroxymethylation, EZH2 expression, the extent of H3K27me3 and EZH2 Tyr 641 somatic mutation in DLBCL patients have not been studied in a comprehensive or integrated way. We aim to see significant impact of DNA methylation, DNA hydroxymethylation, EZH2 expression levels and mutation status, and H3K27 trimethylation on the clinical and biological presentation of DLBCL. Patients and Methods We enrolled 107 consecutive patients with DLBCL in Far Eastern Memorial Hospital and 83 consecutive patients in Chi Mei Medical Center (diagnosed between 2002 and 2009). The demographic data, treatment regimens, and response of disease were reviewed retrospectively. Immunohistochemistry (IHC) was used to examine 5-mC, 5-hmC, EZH2 expression and the extent of H3K27me3 in formalin-fixed, paraffin-embedded biopsy specimens of DLBCL. DNA extraction from the tissues was examined for EZH2 Tyr641 mutation. Statistical analysis was performed with the Stata statistical software (Small Stata, version 11.0, Stata Corp, College Station, TX). Results Statistical analysis was performed in 138 patients with Rituximab-CHOP and CHOP regimen. Totally we recruited 90 male (65%) and 48 female (35%) with a median age 57.2 years. Fifty-nine percent of the patients had stage I/II disease. According to the IPI, 70% of the patients were classified as low/low-intermediate risk (IPI=0-2) and 30% as intermediate-high/high risk (IPI=3-5). Thirty-seven patients received CHOP-like regimen and 101 patients received Rituximab-CHOP-like regimen as first-line chemotherapy. The median observation time for overall survival (OS) in the 138 patients was 42 mo. Male patients tended to have lower H3K27me3 expression (p=0.044). Certain clinical parameters such as IPI score, stage and response to treatment were correlated with OS. None of 48 patients harbored EZH2 mutation at Tyr641. Low expression 5-mC patients tended to have more complete response after standard chemotherapy (P=0.006). Low expression 5-mC tended to have lower H3K27me3 expression (P=0.014). And low expression 5-mC tended to have lower 5-hmC expression (P=0.001). There was no obvious relationship between the expression of EZH2 and degree of H3K27me3 in DLBCL tumor cells. Although there was no significant prognostic impact in univariate survival analysis, low 5-hmC expression seemed to have longer OS (P=0.061). When other strong prognostic factors were included in multivariate analysis, there was no significant prognostic impact in single epigenetic marker. Then we subdivide the patients by combination H3K27me3 with 5-hmC. The high H3K27me3/ low 5-hmC patients was associated with longer OS (P=0.0497) but there was no significant prognostic impact in multivariate survival analysis. In the subgroup analysis, we enrolled the 110 patients with complete remission (CR) after standard chemotherapy and subdivide the patients by combination H3K27me3 with 5-hmC. The high H3K27me3/ low 5-hmC patients was associated with longer OS in univariate (P=0.0131) and multivariate analysis (P=0.017). Conclusion High H3K27me3/ low 5-hmC expression was a possible favorable prognostic factor in DLBCL and it showed even more significant prognostic impact in CR patients. Theincidence of EZH2 mutation at Tyr641 in our cohort is much lower compared with western countries. Low expression 5-mC tended to have lower H3K27me3 expression and low expression 5-mC tended to have lower 5-hmC in DLBCL tumor cells. Our study suggests epigenetic markers may be an informative biomarker for prognosis prediction in DLBCL. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T23:38:19Z (GMT). No. of bitstreams: 1 ntu-101-P99421011-1.pdf: 1672312 bytes, checksum: 843b6da0e7e32758eac7cd92d51360d3 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 口試委員會審定書…………………….……………………………………… ii
誌謝…………………….………………………………………………………. iii 中文摘要………………………………………………………………………. 1 英文摘要………………………………………….……………………………. 4 目錄…………………………………………………………………………… 7 第一章 緒論………………………………………………………………… 11 第一節 癌細胞與表觀基因學的關係……….………………………… 12 第二節 EZH2在細胞中的角色…………………………………… …. 15 第三節 EZH2的突變與淋巴癌的關係……………………………….. 16 第四節 研究假說與特定目的………………………………………….. 17 第二章 研究方法與材料……………………………………………………. 20 第一節 研究設計與研究對象………………………………… ……… 21 第二節 免疫染色(IHC)……………………………………………… 21 第三節 semi-nested PCR與偵測EZH2 Tyr641突變………………. 22 第四節 臨床資料收集………………………………………………. 23 第五節 統計分析………………… ……………………………………. 23 第三章 研究結果……………………………………………………………. 24 第一節 病患臨床資料…………………………………………….…… 25 第二節 EZH2 Tyr641mutation……………………………….………….. 25 第三節 IHC stain與四種抗體之間的關係…. …….…….………… 25 第四節 病患臨床資料與分析結果……………….………………… 26 第四章 討論……………………………………………………….…………. 28 第一節 研究的主要發現………………………………. ……………. 29 第二節 Low 5-hmC的臨床意義與其他癌症的差異性….……………. 29 第三節 Low 5-mC的臨床意義………. ……………………………. 30 第四節 EZH2的臨床意義與其他癌症的差異性….………………. 30 第五節 EZH2與H3K27me3在淋巴癌細胞中的相關性……………. 30 第六節 H3K27me3與5-mC在淋巴癌細胞中的相關性.……………. 31 第七節 5-mC與5-hmC在淋巴癌細胞中的相關性……………….. 31 第八節 EZH2 Tyr641突變在亞洲國家與歐美國家的差異性….…… 32 第九節 研究的限制……………………………………………………. 32 第五章 結論與未來展望……………………… …………………………… 33 第一節 結論……………………………… ……………………….. … 34 第二節 展望………………… …………………………………………. 34 第六章 圖表………………… ……………………………………………… 35 第七章 參考文獻…………………………………………………………….. 69 圖目錄 圖一 四種免疫染色陽性的顯微鏡照片………………………………… 36 圖二 以EZH2_015F3與EZH2_015R所放大的DNA片段……………. 37 圖三 以EZH2_015FF與EZH2_015R所放大的DNA片段…………… 38 圖四 檢體所放大的DNA片段進行定序…………………….………….. 39 圖五 生存曲線分析(年紀)………. …………………………………… 40 圖六 生存曲線分析(治療的處方)……………………………………. 41 圖七 生存曲線分析(治療後的反應)……………. …………………… 42 圖八 生存曲線分析(IPI score)……………………………………. …. 43 圖九 生存曲線分析(Stage)…………………….……………………… 44 圖十 生存曲線分析(EZH2)…………………….……………………… 45 圖十一 生存曲線分析(H3K27me3)……………………………………… 46 圖十二 生存曲線分析(5-mC)……………………………………………. 47 圖十三 生存曲線分析(5-hmC)………………………………………….. 48 圖十四 生存曲線分析(H3K27me3/ 5-hmC)…………………………. 49 圖十五 生存曲線分析(H3K27me3/ 5-mC)……………………..……. 50 圖十六 生存曲線分析(110位經治療後完全緩解的病人IPI score).…. 51 圖十七 生存曲線分析(110位經治療後完全緩解的病人H3K27me3/ 5-hmC的新分類)…………………………………………………. 52 表目錄 表一 EZH2與H3K27me3在不同癌症中預後的狀況………………. …. 53 表二 EZH2與H3K27me3在不同淋巴癌中表現的狀況……………….. 54 表三 EZH2引子(primer)設計…………………………………………….. 55 表四 四種免疫染色的廠牌與設定………………………………………. 56 表五 四種免疫染色的判讀標準…………………………………………. 57 表六 190 位病患的臨床資料……………………………………………. 58 表七 四種免疫染色彼此間的相關分析……. ……………………………. 59 表八 190位病患的臨床資料與四種免疫染色間的相關分析…………. 60 表九 138 位接受CHOP/ R-CHOP的病患用來分析OS的臨床資料…. 61 表十 138 位接受CHOP/ R-CHOP病患的臨床資料與四種免疫染色 的相關分析………………………………………………………. 62 表十一 138 位接受CHOP/ R-CHOP病患的臨床資料與四種免疫染色的 生存分析(單變項)……………………………………………. 63 表十二 138 位接受CHOP/ R-CHOP病患的臨床資料和5-hmC的生存 分析(多變項)…………………………………………………. 64 表十三 138 位接受CHOP/ R-CHOP以H3K27me3/ 5-hmC的新分類之 臨床資料和相關分析……………………………………………. 65 表十四 138 位接受CHOP/ R-CHOP病患的臨床資料和H3K27me3/ 5-hmC新分類的生存分析(多變項)……………………………. 66 表十五 110位經治療後完全緩解的病人之臨床資料和四種免疫染色的 生存分析(單變項)…………………………………………..……. 67 表十六 110位經治療後完全緩解病患的臨床資料和H3K27me3/ 5-hmC的 生存分析(多變項)………………………………………………. 68 | |
dc.language.iso | zh-TW | |
dc.title | EZH2的表現、H3K27三個甲基化與DNA甲基化在瀰漫性大型B細胞淋巴癌預後的影響 | zh_TW |
dc.title | Prognostic impact of EZH2 expression, H3K27 trimethylation and DNA methylation in diffuse large B cell lymphoma | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林亮音,葉坤輝 | |
dc.subject.keyword | 淋巴癌,表觀基因學,甲基化與組蛋白, | zh_TW |
dc.subject.keyword | Diffuse large B cell lymphoma,epigenetics,EZH2,H3H27me3,5-methylcytosine,5-hydroxymethylcytosine, | en |
dc.relation.page | 74 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-07-26 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 臨床醫學研究所 | zh_TW |
顯示於系所單位: | 臨床醫學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-101-1.pdf 目前未授權公開取用 | 1.63 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。