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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林欽塘 | |
dc.contributor.author | Hsin-Lan Su | en |
dc.contributor.author | 蘇欣蘭 | zh_TW |
dc.date.accessioned | 2021-06-08T02:52:02Z | - |
dc.date.copyright | 2017-09-13 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-08-14 | |
dc.identifier.citation | Adam, F. (2005) microRNA : A Revolution in Gene Expression. Journal of Young Investigators.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/20530 | - |
dc.description.abstract | 相較於其他癌症,鼻咽癌具有相當高的「地區性」,盛行於中國大陸東南沿海、香港、新加坡、台灣等地,其致病機轉相當複雜。雖然診斷及治療方法不斷進步,但因其發病部位較為隱密,待病患查覺異狀至確診,往往已是晚期,使得預後不良。近期有研究指出Epstein-Barr virus (EBV)與鼻咽癌的惡化有關。在本次研究中發現鼻咽癌細胞株經異體移植(xenografts)得到的同源亞細胞株(sub-cell line),其腫瘤生長力(tumorgenesis)遠大於原細胞株。分析兩細胞株的表現和細胞行為,可得知鼻咽癌亞細胞株在細胞增殖(proliferation)及細胞侵犯(invasion)上具有較強勢的作用。已知microRNA可調控細胞的基因表現,比較兩種細胞株發現在亞細胞株中之hsa-miR-486表現量明顯較高。經由RNA微陣列分析(RNA microarray analysis)發現Reversion-inducing-cysteine-rich protein with kazal motifs (RECK)腫瘤抑制基因會受到miR-486的調節。我們找到RECK基因上miR-486的結合位,將建構好具有miR-486結合位的RECK基因mRNA片斷載體轉染入此二細胞株中,以觀察其對腫瘤發生的能力,並同時進行老鼠活體試驗,從而證實miR-486可抑制RECK基因造成鼻咽癌細胞的腫瘤發生。RECK基因可調控鼻咽癌之生長在之前的SXO-5基因表達之研究中已發現,但與miR-486之關係在鼻咽癌的進展中未曾被發現過,在未來研發中可預期其可能作為分子標靶之治療。 | zh_TW |
dc.description.abstract | To compare with other cancer disease, nasopharyngeal carcinoma (NPC) often occurs in local region, such as in the southeast coast of the mainland China , Hong Kong, Singapore , and Taiwan. Its pathogenesis is quite complex. Despite advances in diagnosis and treatment , and because of the incidence of the more intimate parts , the abnormal symptoms in patients with perceptible to the diagnosis often is late, resulting in a poor prognosis . Recent researches have pointed out that Epstein-Barr virus (EBV) is one of the reasons that cause nasopharyngeal carcinoma progression. In our laboratory study, we have established a NPC sub-line from one NPC line which was produced from the NPC xenografts. This sub-line showed a higher capacity to induce tumor progression. Analysis of the behavior and phenotype of the original and sub-line showed that NPC sub-line had more higher potential to proliferation, and invasion. Since microRNA can regulate gene expression, we found that the expression of miR-486 has significantly higher in the sub-line. Via RNA microarray analysis of over expressed miR-486, we found that reversion-inducing-cysteine-rich protein with kazal motifs (RECK) gene (a metastasis suppressor) was down regulated by miR-486. Since we have identified that miR-486 binding site presented in RECK gene, therefor we constructed the specific vector expression to transfect in those two cell line. Results showed that miR-486 can regulate RECK gene in two transfected cell lines. At the same time, we have also confirmed the identical results in the in vivo experiment. This is a new finding that RECK gene can regulate NPC progression through miR-486 transfection. In the future for development of NPC treatment, miR-486 may be used as a molecular targeted therapy. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T02:52:02Z (GMT). No. of bitstreams: 1 ntu-106-R00444007-1.pdf: 2724179 bytes, checksum: 65beec36bc3e2e3d324e71a2507a3554 (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | 目錄 Table of Contents
口試委員審定書 i 致謝 ii 中文摘要 iii Abstract iv List of Figures viii Introduction 1 1. Nasopharyngeal carcinoma (NPC) 1 2. Etiology of NPC 4 3. microRNA 6 4. miRNA-486 11 5. RECK gene 13 Materials and Methods 17 1. Cell lines 17 2. miRNA 17 3. Extraction of RNA and preparation of cDNA 18 4. Quantitative Time-Lapse Microscopy images 19 5. cDNA Microarray assay 20 6. MTT Assay 21 7. Scratch wound healing assay 21 8. Amplification and Extraction of plasmids 22 9. Mini-plasmid purification 22 10. Invasion assay 23 11. Luciferase assay 24 12. Western Blotting 25 13. Immunohistochemical staining 26 14. In vivo assay of xenograft growth 27 15. Statistical analysis 28 Results 29 1. Cell morphology 29 2. miR-486 regulated NPC cell behavior 29 3. RECK was the miR-486 target gene 30 4. RECK expression was higher in NPC-TW01 than in NPC-TW01N1 cell line 31 5. Immunostaining of RECK protein in NPC biopsy specimens 31 6. Functional analysis of miR-486 in SCID mice bearing NPC xenografts 31 Discussion 33 References 36 Tables 60 List of Figures Figure 1. Cell Morphology 47 Figure 2. Reduction of cell migration ability in miRNA transfected NPC cell were shown. 48 Figure 3. Differences in proliferation between NPC-TW01 and NPC-TW01N1 ( A) Quantitative Time-Lapse Microscopy images. (B) MTT assay 49 Figure 4. Differences in proliferation between NPC-TW01 transfected miR-486 mimic and negative control ( A) Quantitative Time-Lapse Microscopy images. (B) MTT assay 50 Figure 5. Differences in proliferation between NPC-TW01N1 transfected miR-486 inhibitor and negative control ( A) Quantitative Time-Lapse Microscopy images. (B) MTT assay 51 Figure 6. Differences in invasion ability between NPC-TW01 and NPC-TW01N1 52 Figure 7. Invasion ability is up regulation by miR-486 mimic in NPC-TW01 53 Figure 8. Invasion ability is down regulation by miR-486 inhibitor in NPC-TW01N1 54 Figure 9. Luciferase assay confirm the miR-486 target gene is RECK 55 Figure 10. The expression of RECK protein in NPC-TW01 and NPC-TW01N1 cells. 56 Figure 11. 20 NPC patient’s biopsy specimens were examined by RECK immunostaining. 57 Figure 12. SCID mice bearing NPC-TW01N1 cells after the latter were transfected with miR-486 inhibitor showing marked decrease of xenograft sizes. 58 | |
dc.language.iso | en | |
dc.title | miR-486對鼻咽癌的發生所扮演之功能角色 | zh_TW |
dc.title | The Functional Role of miR-486 in Nasopharyngeal Carcinoma Tumorigenesis | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 楊雅倩,張鑫,劉雅雯,吳漢忠 | |
dc.subject.keyword | 鼻咽癌,RECK基因,miR-486,細胞增殖,細胞侵犯, | zh_TW |
dc.subject.keyword | NPC tumorigenesis,miR-486,reversion-inducing-cysteine-rich protein with kazal motifs (RECK),metastasis suppressor, | en |
dc.relation.page | 60 | |
dc.identifier.doi | 10.6342/NTU201703116 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2017-08-14 | |
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 |
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