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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 賴亮全(Liang-Chuan Lai) | |
dc.contributor.author | Li-Ju Wang | en |
dc.contributor.author | 王麗茹 | zh_TW |
dc.date.accessioned | 2021-06-16T08:05:56Z | - |
dc.date.available | 2019-10-09 | |
dc.date.copyright | 2014-10-09 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2014-06-23 | |
dc.identifier.citation | 1. Nathoo, N., et al., Pathobiology of brain metastases. J Clin Pathol, 2005. 58(3): p. 237-42.
2. Hoffman PC, M.A., Vokes EE., Lung cancer. Lancet, 2000: p. 479-85. 3. Department of Health, Executive Yuan, R.O.C. 2013; Available from: http://www.doh.gov.tw/CHT2006/DM/DM2_p01.aspx?class_no=25&level_no=1&doc_no=88618. 4. Thun, M.J., et al., Lung cancer occurrence in never-smokers: an analysis of 13 cohorts and 22 cancer registry studies. PLoS Med, 2008. 5(9): p. e185. 5. Bunn, P.A., Jr. and K. Kelly, New chemotherapeutic agents prolong survival and improve quality of life in non-small cell lung cancer: a review of the literature and future directions. Clin Cancer Res, 1998. 4(5): p. 1087-100. 6. Grinberg-Rashi, H., et al., The expression of three genes in primary non-small cell lung cancer is associated with metastatic spread to the brain. Clin Cancer Res, 2009. 15(5): p. 1755-61. 7. Hayama, M., et al., One-step Nucleic Acid Amplification for Detection of Lymph Node Metastasis in Lung Cancer. Ann Thorac Cardiovasc Surg, 2013. 8. Coleman, R.E., Clinical features of metastatic bone disease and risk of skeletal morbidity. Clin Cancer Res, 2006. 12(20 Pt 2): p. 6243s-6249s. 9. Slotman, B., et al., Prophylactic cranial irradiation in extensive small-cell lung cancer. N Engl J Med, 2007. 357(7): p. 664-72. 10. Meert, A.P., et al., Prophylactic cranial irradiation in small cell lung cancer: a systematic review of the literature with meta-analysis. BMC Cancer, 2001. 1: p. 5. 11. Lee, J.M., et al., The epithelial-mesenchymal transition: new insights in signaling, development, and disease. J Cell Biol, 2006. 172(7): p. 973-81. 12. Hay, E.D., The mesenchymal cell, its role in the embryo, and the remarkable signaling mechanisms that create it. Dev Dyn, 2005. 233(3): p. 706-20. 13. Kalluri, R. and R.A. Weinberg, The basics of epithelial-mesenchymal transition. J Clin Invest, 2009. 119(6): p. 1420-8. 14. Ferrara, N., H.P. Gerber, and J. LeCouter, The biology of VEGF and its receptors. Nat Med, 2003. 9(6): p. 669-76. 15. Walsh, F.S., et al., N-cadherin gene maps to human chromosome 18 and is not linked to the E-cadherin gene. J Neurochem, 1990. 55(3): p. 805-12. 16. Reid, R.A. and J.J. Hemperly, Human N-cadherin: nucleotide and deduced amino acid sequence. Nucleic Acids Res, 1990. 18(19): p. 5896. 17. Hulit, J., et al., N-cadherin signaling potentiates mammary tumor metastasis via enhanced extracellular signal-regulated kinase activation. Cancer Res, 2007. 67(7): p. 3106-16. 18. Gravdal, K., et al., A switch from E-cadherin to N-cadherin expression indicates epithelial to mesenchymal transition and is of strong and independent importance for the progress of prostate cancer. Clin Cancer Res, 2007. 13(23): p. 7003-11. 19. Nieman, M.T., et al., N-cadherin promotes motility in human breast cancer cells regardless of their E-cadherin expression. J Cell Biol, 1999. 147(3): p. 631-44. 20. Ding, J., et al., [Expression and significance of Slug, E-cadherin and N-cadherin in gastrointestinal stromal tumors]. Zhonghua Yi Xue Za Zhi, 2012. 92(4): p. 264-8. 21. Tomita, K., et al., Cadherin switching in human prostate cancer progression. Cancer Res, 2000. 60(13): p. 3650-4. 22. Su, H., et al., Immediate expression of Cdh2 is essential for efficient neural differentiation of mouse induced pluripotent stem cells. Stem Cell Res, 2013. 10(3): p. 338-48. 23. Marie, P.J., et al., Cadherin-Mediated Cell-Cell Adhesion and Signaling in the Skeleton. Calcif Tissue Int, 2013. 24. Yamashita, S., et al., Chemical genomic screening for methylation-silenced genes in gastric cancer cell lines using 5-aza-2'-deoxycytidine treatment and oligonucleotide microarray. Cancer Sci, 2006. 97(1): p. 64-71. 25. Alexander, N.R., et al., N-cadherin gene expression in prostate carcinoma is modulated by integrin-dependent nuclear translocation of Twist1. Cancer Res, 2006. 66(7): p. 3365-9. 26. Rosivatz, E., et al., Differential expression of the epithelial-mesenchymal transition regulators snail, SIP1, and twist in gastric cancer. Am J Pathol, 2002. 161(5): p. 1881-91. 27. Ng, Y.H., H. Zhu, and P.C. Leung, Twist modulates human trophoblastic cell invasion via regulation of N-cadherin. Endocrinology, 2012. 153(2): p. 925-36. 28. Barbieri, C.E., et al., Loss of p63 leads to increased cell migration and up-regulation of genes involved in invasion and metastasis. Cancer Res, 2006. 66(15): p. 7589-97. 29. Dwyer, M.A., et al., WNT11 expression is induced by estrogen-related receptor alpha and beta-catenin and acts in an autocrine manner to increase cancer cell migration. Cancer Res, 2010. 70(22): p. 9298-308. 30. Lewis, B.P., C.B. Burge, and D.P. Bartel, Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell, 2005. 120(1): p. 15-20. 31. Brennecke, J., et al., Principles of microRNA-target recognition. PLoS Biol, 2005. 3(3): p. e85. 32. Xie, X., et al., Systematic discovery of regulatory motifs in human promoters and 3' UTRs by comparison of several mammals. Nature, 2005. 434(7031): p. 338-45. 33. Grun, D., et al., microRNA target predictions across seven Drosophila species and comparison to mammalian targets. PLoS Comput Biol, 2005. 1(1): p. e13. 34. Gao, P., et al., The molecular mechanism of microRNA-145 to suppress invasion-metastasis cascade in gastric cancer. Oncogene, 2013. 32(4): p. 491-501. 35. Lee, R.C., R.L. Feinbaum, and V. Ambros, The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell, 1993. 75(5): p. 843-54. 36. Monteys, A.M., et al., Structure and activity of putative intronic miRNA promoters. RNA, 2010. 16(3): p. 495-505. 37. Shi, X.B., C.G. Tepper, and R.W. deVere White, Cancerous miRNAs and their regulation. Cell Cycle, 2008. 7(11): p. 1529-38. 38. Clement, J.Q., et al., The stability and fate of a spliced intron from vertebrate cells. RNA, 1999. 5(2): p. 206-20. 39. Lee, Y., et al., MicroRNA genes are transcribed by RNA polymerase II. EMBO J, 2004. 23(20): p. 4051-60. 40. Tomari, Y. and P.D. Zamore, Perspective: machines for RNAi. Genes Dev, 2005. 19(5): p. 517-29. 41. Winter, J., et al., Many roads to maturity: microRNA biogenesis pathways and their regulation. Nat Cell Biol, 2009. 11(3): p. 228-34. 42. Tie, J., et al., MiR-218 inhibits invasion and metastasis of gastric cancer by targeting the Robo1 receptor. PLoS Genet, 2010. 6(3): p. e1000879. 43. Denk, A.E., et al., Slit3 inhibits activator protein 1-mediated migration of malignant melanoma cells. Int J Mol Med, 2011. 28(5): p. 721-6. 44. Guan, H., et al., Down-regulation of miR-218-2 and its host gene SLIT3 cooperate to promote invasion and progression of thyroid cancer. J Clin Endocrinol Metab, 2013. 45. Marlow, R., et al., SLITs suppress tumor growth in vivo by silencing Sdf1/Cxcr4 within breast epithelium. Cancer Res, 2008. 68(19): p. 7819-27. 46. Tseng, R.C., et al., SLIT2 attenuation during lung cancer progression deregulates beta-catenin and E-cadherin and associates with poor prognosis. Cancer Res, 2010. 70(2): p. 543-51. 47. Alajez, N.M., et al., MiR-218 suppresses nasopharyngeal cancer progression through downregulation of survivin and the SLIT2-ROBO1 pathway. Cancer Res, 2011. 71(6): p. 2381-91. 48. Uesugi, A., et al., The tumor suppressive microRNA miR-218 targets the mTOR component Rictor and inhibits AKT phosphorylation in oral cancer. Cancer Res, 2011. 71(17): p. 5765-78. 49. Kinoshita, T., et al., Tumor suppressive microRNA-218 inhibits cancer cell migration and invasion through targeting laminin-332 in head and neck squamous cell carcinoma. Oncotarget, 2012. 3(11): p. 1386-400. 50. Liu, Y., et al., MiR-218 reverses high invasiveness of glioblastoma cells by targeting the oncogenic transcription factor LEF1. Oncol Rep, 2012. 28(3): p. 1013-21. 51. Goossens, K., et al., Regulatory microRNA Network Identification in Bovine Blastocyst Development. Stem Cells Dev, 2013. 22(13): p. 1907-20. 52. Hassan, M.Q., et al., miR-218 directs a Wnt signaling circuit to promote differentiation of osteoblasts and osteomimicry of metastatic cancer cells. J Biol Chem, 2012. 287(50): p. 42084-92. 53. Martini, N., et al., Incidence of local recurrence and second primary tumors in resected stage I lung cancer. J Thorac Cardiovasc Surg, 1995. 109(1): p. 120-9. 54. Huang, Q. and X. Ouyang, Predictive biochemical-markers for the development of brain metastases from lung cancer: Clinical evidence and future directions. Cancer Epidemiol, 2013. 55. Li, B.S., et al., Plasma microRNAs, miR-223, miR-21 and miR-218, as novel potential biomarkers for gastric cancer detection. PLoS One, 2012. 7(7): p. e41629. 56. Dallol, A., et al., SLIT2, a human homologue of the Drosophila Slit2 gene, has tumor suppressor activity and is frequently inactivated in lung and breast cancers. Cancer Res, 2002. 62(20): p. 5874-80. 57. Davidson, M.R., et al., MicroRNA-218 is deleted and downregulated in lung squamous cell carcinoma. PLoS One, 2010. 5(9): p. e12560. 58. Chen, W.F., et al., SLIT2 inhibits cell migration in colorectal cancer through the AKT-GSK3beta signaling pathway. Int J Colorectal Dis, 2013. 28(7): p. 933-40. 59. Bartel, D.P., MicroRNAs: genomics, biogenesis, mechanism, and function. Cell, 2004. 116(2): p. 281-97. 60. Lu, T.P., et al., miRSystem: an integrated system for characterizing enriched functions and pathways of microRNA targets. PLoS One, 2012. 7(8): p. e42390. 61. Shi, K., et al., MicroRNA-214 suppresses osteogenic differentiation of C2C12 myoblast cells by targeting Osterix. Bone, 2013. 55(2): p. 487-94. 62. Wu, D.W., et al., Paxillin predicts survival and relapse in non-small cell lung cancer by microRNA-218 targeting. Cancer Res, 2010. 70(24): p. 10392-401. 63. Sahai, E., Mechanisms of cancer cell invasion. Current Opinion in Genetics & Development, 2005. 15(1): p. 87-96. 64. Mehlen, P. and A. Puisieux, Metastasis: a question of life or death. Nat Rev Cancer, 2006. 6(6): p. 449-58. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58099 | - |
dc.description.abstract | 肺癌為造成癌症死亡中最主要的癌症之一,而大腦為肺癌細胞主要轉移的器官。目前臨床上,主要治療腦轉移的方式為放射性治療,但由於放射性治療有很嚴重的副作用,找尋新的且有效率的腦轉移治療方式已成為當前研究中備受關注的議題。目前眾多的實驗治療方法之一,為使用微小 RNA (MicroRNA)去抑制具有腦轉移能力的肺癌細胞,但此領域的發展與對分子層次調控機制的所知相當有限。所以本篇研究中我們將尋找可以抑制肺腺癌細胞轉移的miRNA,並對其分子調控進行探討。
在本篇研究中,我們將使用一具有腦轉移能力的肺癌細胞株─ BM#7 ─作為本篇實驗的研究對象。由於CDH2的過度表現已被證實為細胞轉移的指標,我們利用Illumina miRNA微陣列晶片(microarray)去篩選BM#7細胞與 F4細胞(不具有腦轉移能力的肺癌細胞)相比表現量有差異的miRNA。並利用生物資訊工具去預測並挑選這一群有差異的miRNA中與CDH2之間有鍵結關係的miRNA,最後我們挑選出九個miRNA。由於CDH2在BM#7中表現量上升,我們藉由各種不同的預測方式挑選出與CDH2親和力最高且與CDH2有相反表現的miRNA─ miR-218。我們進一步設計實驗驗證:miR-218的異常表現上升,會促使CDH2表現量下降並抑制肺癌細胞的轉移。 我們先利用即時定量聚合酶連鎖反應 (real-time PCR)去驗證內生性CDH2和miR-218的表現量。結果顯示,在BM#7中miR-218的表現量是下降的,為了瞭解究竟是何種機制促使miR-218下降,因此我們去偵測miR-218的前驅物。由於miR-218的前驅物有兩個分別存在於兩個基因:SLIT2 (pri-mir-218-1)和SLIT3 (pri-mir-218-2)。根據real-time PCR結果表示,miR-218的表現量下降是因為pri-mir-218-1所導致的。之後我們想要再去驗證miR-218是否會與CDH2-3’UTR鍵結,因此我們利用冷光酵素活性測定(Luciferase assay)來證明。結果顯示,miR-218在CDH2-3’UTR的有兩個鍵結位置。最後我們利用細胞穿透試驗 (transwell assay)來探討BM#7細胞中大量表現miR-218在CDH2上的功能影響。由結果看來,大量表現miR-218會抑制BM#7細胞的轉移能力。 綜合以上所述,本篇研究顯示在BM#7中miR-218表現量下降是因為pri-mir-218-1表現量降低所致。此外也證明miR-218可以藉由與CDH2-3'UTR鍵結抑制CDH2表現,進而使細胞轉移能力下降。故miR-218和SLIT2在肺癌細胞腦轉移的臨床治療中,可能可以做為一個新的治療方式與指標。 | zh_TW |
dc.description.abstract | Lung cancer is the leading cause of cancer-related mortality in the world. Brain is a major migratory site of lung cancer[1]. So far, the major therapy of brain metastasis is irradiation, but it has a serious side effect. Hence, there is a great need to find efficiency methods for curing brain metastasis. One possible therapeutic strategy is using miRNA to inhibit brain metastatic lung cancer. However, the knowledge of its regulatory mechanism is still very limited. Therefore, as an initial step, the purpose of this study is trying to identify miRNAs that can inhibit the migration of lung adenocarcinoma cells.
Here, a brain metastatic lung adenocarcinoma cell line, BM#7, was used as an experimental model. The expression of CDH2 was used as a biomarker for representing metastatic ability. Illumina miRNA microarrays were used to screen the differentially expressed miRNAs in BM#7 cells and parental F4 cells. Bioinformatic tools were used to predict which miRNA can target to CDH2. We identified nine miRNAs that differentially expressed in BM#7 and predicted to target to CDH2. Among these nine miRNAs, since CDH2 was up-regulated in BM#7, we focused on miR-218 that was down-regulated and predicted to target CDH2 by several algorithms. The endogenous expression levels of CDH2 and miR-218 were validated by real-time PCR. Next, to understand the regulatory mechanism of miR-218, we examined the expression levels of miR-218 precursors. Since miR-218 had two precursors, pri-mir-218-1 and pri-mir-218-2, located in SLIT2 and SLIT3 respectively, real-time PCR was used to measure their expression levels. The results showed that decline of miR-218 in BM#7 were due to down-regulation of pri-mir-218-1. Third, to explore whether miR-218 could bind to CDH2, luciferase assays were conducted. The results showed that miR-218 could directly bind to CDH2-3’UTR at two binding sites. Lastly, to investigate the function role of miR-218 on CDH2, transwell migration assay was conducted in BM#7 with miR-218 over-expression. The ability of cell migration was suppressed when miR-218 was up-regulated in BM#7 cells. Taken together, this study showed that the down-regulation of miR-218 was attributed to the decreased expression of pri-mir-218-1 in BM#7, and that miR-218 could inhibit cell migration by targeting to CDH2-3’UTR. MiR-218 might be a novel drug for developing clinical therapies for lung cancer with brain metastasis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T08:05:56Z (GMT). No. of bitstreams: 1 ntu-102-R00441005-1.pdf: 1616674 bytes, checksum: 2f7d7038e943898f80218f5688772e2f (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | Chapter 1 Introduction 1
1.1 Lung cancer and metastasis 1 1.2 CDH2 is a epithelial- mesenchymal transition marker 2 1.3 MicroRNA and miR-218 4 1.4 Motivation and Specific Aims 5 Chapter 2 Materials and Methods 7 2.1 Cell culture 7 2.2 Construct design and cell transfections 7 2.3 Luciferase reporter assay 9 2.4 Western blot 10 2.5 Reverse transcription and quantitative Real-Time PCR 10 2.6 Microarray data analysis 12 2.7 Cell migration assay 13 Chapter 3 Results 15 3.1 CDH2 was up-regulated in metastatic cell lines. 15 3.2 Identification of miRNAs by array-based hybridization 17 3.3 Pri-mir-218-1 and its original gene SLIT2 were down-regulated in metastatic cells. 20 3.4 MiR-218 could directly bind to CDH2-3’UTR. 23 3.5 Over-expression of miR-218 could suppress the expression of CDH2. 24 3.6 Over-expression of miR-218 could suppress cell migration by down-regulating the expression of CDH2. 27 Chapter 4 Discussion 30 4.1 MiR-218 was expressed from pri-mir-218-1, not pri-mir-218-2, in lung cancer with brain metastasis. 30 4.2 MiR-218 can target two binding sites in the 3’UTR of CDH2. 32 4.3 Limitations and future study 33 4.4 Summary 34 REFERENCES 36 | |
dc.language.iso | en | |
dc.title | 來自於pri-mir-218-1的miR-218可藉由調控N-Cadherin來抑制肺腺癌細胞的移動 | zh_TW |
dc.title | MiR-218, Expressed From Pri-mir-218-1, Inhibits Migration of Lung Adenocarcinoma Cells by Suppressing N-Cadherin | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 佘玉萍(Yuh-Pyng Sher) | |
dc.contributor.oralexamcommittee | 莊曜宇(Eric Y. Chuang),蔡孟勳(Mon-Hsun Tsai) | |
dc.subject.keyword | 微小RNA,miR-218,肺癌,腦轉移,SLIT2, | zh_TW |
dc.subject.keyword | MicroRNA,miR-218,SLIT2,metastasis,lung adenocarcinoma, | en |
dc.relation.page | 40 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2014-06-24 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 生理學研究所 | zh_TW |
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