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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56976完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 廖泰慶(Albert Taiching Liao) | |
| dc.contributor.author | Rui-Hong Wu | en |
| dc.contributor.author | 吳瑞紘 | zh_TW |
| dc.date.accessioned | 2021-06-16T06:32:15Z | - |
| dc.date.available | 2016-08-08 | |
| dc.date.copyright | 2014-08-08 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-08-05 | |
| dc.identifier.citation | Aalipour, A., Advani, R.H., 2013. Bruton tyrosine kinase inhibitors: a promising novel targeted treatment for B cell lymphomas. British journal of haematology 163, 436-443.
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Zwingenberger, A.L., Vernau, W., Shi, C., Yan, W., Chen, X., Gordon, I.K., Kent, M.S., 2012. Development and characterization of 5 canine B-cell lymphoma cell lines. Leukemia research 36, 601-606. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56976 | - |
| dc.description.abstract | 淋巴瘤是犬隻最常見的造血器官腫瘤，在臨床上通常會以多種藥劑併用的化療方法來進行治療，而接受治療的犬隻，在治療初期大都會有良好的反應，大約可以使80-90 ％患犬的淋巴瘤消退。然而，大多數的犬隻最終還是會由於抗藥性的產生而復發，平均約發生於診斷後一年。由於現今沒有比化療更有效用的治療方法，因此在過去的治療上，犬淋巴瘤分類僅簡單的區分為B細胞或T細胞兩型。因此，在治療犬隻淋巴瘤上，必須為犬淋巴瘤進行更多的分類且提供不同的治療藥物。在這項研究中為了確定犬淋巴瘤細胞株 CLBL -1（B細胞型），CLC (非T非B細胞型)和UL-1（T細胞型）的表現型，我們首先使用了流式細胞儀偵測了CD3、CD4、CD5、CD8、CD21、CD34、CD45、CD56和CD79αcy 抗體表現，用免疫細胞化學染色法偵測了MUM -1、Bcl-6、Bcl-2、CD10和KMO的蛋白表現，並以PCR / RT-PCR法偵測了p53、FLT3、NRAS和BRAF基因有無突變。而在UL-1細胞株，我們發現在p53基因有一個G485A的點突變，造成了R162H突變。接著我們以小分子抑製劑， Kynurenine 3-monooxygenase (KMO)抑製劑和新型烷基化藥物BO-1055、BO-1922、BO-2094和BO-1978來進行細胞增生試驗，其結果發現此三種類型的藥物都能顯著降低三種細胞株的增殖並造成細胞死亡。此實驗結果表示這些藥物俱有治療犬隻淋巴瘤的潛力，而未來的研究將集中在藥物作用機制的分析。 | zh_TW |
| dc.description.abstract | Canine lymphoma is the most common hematopoietic tumor affecting dog. The remission rate of canine lymphoma treated by available combination chemotherapy protocols is usually high (80%-90%). However, most dogs eventually succumb to drug-resistant recurrence, on average, 1 year after diagnosis. Since no more therapeutic option other than chemotherapy, canine lymphoma is simply classified as B cell or T cell type so far. Therefore, offering more therapeutic options for different types of canine lymphoma will be necessary. In this study, flow cytometry (CD3, CD4, CD5, CD8, CD21, CD34, CD45, CD56, and CD79αcy), immunocytochemistry (MUM-1, Bcl-6, Bcl-2, CD10 and KMO) and PCR/RT-PCR (p53, FLT3, NRAS and BRAF) were used to identify the phenotypes of canine lymphoma cell lines, CLBL-1 (B cell type), CLC (non-T non-B cell type) and UL-1 (T cell type). A G485A point mutation causing R162H mutation was found in p53 gene of UL-1 cells. In subsequent viability assay, small molecule inhibitor, kyneurenine 3-monooxygenase (KMO) inhibitor and novel alkylating agents, BO-1055, BO-1922, BO-2094 and BO-1978, significantly reduced proliferation of all cell lines. This revealed their therapeutic potential on canine lymphoma. Future studies will focus on analysis of drugs mechanism. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T06:32:15Z (GMT). No. of bitstreams: 1 ntu-103-R01629004-1.pdf: 74287905 bytes, checksum: 195e34babae202152bd009b0c103d9cc (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 口試委員審定書 I
誌謝 II 中文摘要 II Abstract IV Contents V Chapter 1. Background and Literature Review 1 1.1 Canine lymphoma 1 1.1.1 Prevalence of canine lymphoma 1 1.1.2 Classification, staging and phenotyping of canine lymphoma 2 1.1.3 Therapy for canine lymphoma 3 1.2 Development of novel therapy for lymphoma 5 1.2.1 Targeted therapy for human lymphoma 5 1.2.2 Tyrosine kinase inhibitor (TKI) 6 1.2.3 Alkylating agents 7 1.2.4 Kynurenine 3-monooxygenase (KMO) 8 1.3 Biomarkers of canine lymphoma 9 1.3.1 Mutations in p53 gene 9 1.3.2 Mutations in flt3 gene 10 1.3.3 Bcl-2, Bcl-6, MUM-1, CD10 11 Chapter 2. Introduction 12 Chapter 3. Materials and methods 15 3.1 Establish the canine lymphoma cell lines 15 3.2 Cell lines and reagents 16 3.3 Cell surface marker analysis 17 3.4 PCR for antigen receptor rearrangement (PARR) 17 3.5 Colony formation assay 18 3.6 Soft agar immunocytochemistry (SA-ICC) 18 3.7 Reverse transcription polymerase chain reaction (RT-PCR) 20 3.8 Examination lymphoma mutation genes 20 3.9 Proliferation assay 22 3.10 Western blotting 23 3.11 Cell cycle analysis 24 3.12 Statistical analysis 25 Chapter 4. Results 26 4.1 Morphology and soft agar colony formation assay in canine lymphoma cells 26 4.2 Canine lymphoma cell line phenotyping 26 4.2.1 Flow cytometry 26 4.2.2 PCR for antigen receptor rearrangement (PARR) 28 4.2.3 Immunocytochemistry (ICC) 28 4.2.4 Polymerase chain reaction (PCR) 28 4.2.5 Reverse transcription polymerase chain reaction (RT-PCR) 29 4.2.6 KMO expression of canine lymphoma cells 29 4.3 Alkylating agents inhibited proliferation of CLBL-1, CLC and UL-1 cells 30 4.4 Dovitinib inhibited proliferation of CLBL-1, CLC and UL-1 cells 31 4.5 KMO inhibitor inhibited proliferation of CLBL-1, CLC and UL-1 cells 32 Chapter 5. Discussion 33 Tables 42 Table 1. World Health Organization clinical staging for domestic animals with lymphoma 42 Table 2. Monoclonal antibodies used for flow cytometry 43 Table 3. Sequence of primers used in PARR 44 Table 4. Sequence of primers used in PCR and RT-PCR 45 Table 5. Sequence of primers used in p53 RT-PCR 46 Table 6. Mutation in p53 gene detected in canine lymphoma cells 47 Table 7. Canine cell line CLBL-1, CLC, UL-1, CLN and DH82 phenotyping results 48 Table 8. Sensitivity of canine lymphoma cells to therapeutic drugs 50 Figures 51 Figure 1. Relative distribution of canine lymphoma subtypes 51 Figure 2. Morphology and soft agar colony formation assay in canine lymphoma cells 52 Figure 3. Comparative the canine lymphoma cell lines CLBL-1, CLC and UL-1 labeled with monoclonal antibodies against canine antigens 54 Figure 4. Comparative the canine lymphoma cell lines CLN large groupn 57 Figure 5. PCR for antigen receptor rearrangement 58 Figure 6. The protein expression of MUM-1, CD10, Bcl-2 and Bcl-6 in canine lymphoma cells by immunocytochemistry 60 Figure 7. PCR and RT-PCR for FLT3 14/15&20, NRAS&BRAF and p53 gene mutations in canine lymphoma cell line 61 Figure 8. The mRNA and protein expression of KMO in canine lymphoma cells 62 Figure 9. Effect of alkylating agents on the cell viability of canine lymphoma cells 65 Figure 10. Effect of dovitinib and KMO inhibitor on the cell viability of canine lymphoma cells 68 Figure 11. Effect of dovitinib on the cell cycle in canine lymphoma cells 70 Figure 12. Effect of KMO inhibitor on the cell cycle in canine lymphoma cells 72 Reference 73 | |
| dc.language.iso | zh-TW | |
| dc.subject | 表型分析 | zh_TW |
| dc.subject | 烷基化藥物 | zh_TW |
| dc.subject | Kynurenine 3-monooxygenase (KMO) | zh_TW |
| dc.subject | dovitinib | zh_TW |
| dc.subject | 流式細胞技術 | zh_TW |
| dc.subject | 犬淋巴瘤 | zh_TW |
| dc.subject | dovitinib | en |
| dc.subject | phenotyping | en |
| dc.subject | flow cytometry | en |
| dc.subject | alkylating agents | en |
| dc.subject | kyneurenine 3-monooxygenase | en |
| dc.subject | Canine lymphoma | en |
| dc.title | 犬淋巴瘤細胞表現型鑑定以及治療藥物之評估 | zh_TW |
| dc.title | Identification of Phenotype and Evaluation of Therapeutic Drugs on Canine Lymphoma Cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林辰栖,李繼忠,張仕杰 | |
| dc.subject.keyword | 犬淋巴瘤,表型分析,流式細胞技術,烷基化藥物,Kynurenine 3-monooxygenase (KMO),dovitinib, | zh_TW |
| dc.subject.keyword | Canine lymphoma,phenotyping,flow cytometry,alkylating agents,kyneurenine 3-monooxygenase,dovitinib, | en |
| dc.relation.page | 86 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2014-08-06 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
| 顯示於系所單位： | 獸醫學系 | |
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