請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17282完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 廖泰慶 | |
| dc.contributor.author | Mei-Hsien Chang | en |
| dc.contributor.author | 張美仙 | zh_TW |
| dc.date.accessioned | 2021-06-08T00:04:47Z | - |
| dc.date.copyright | 2013-08-28 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-08-14 | |
| dc.identifier.citation | Reference
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17282 | - |
| dc.description.abstract | Kynurenine 3-monooxygenase (KMO)是Tryptophan代謝過程中kynurenine pathway中的一個酵素，位於粒腺體的外膜，可將L-kynurenine降解為3-hydroxy kynurenine，而kynurenine pathway的終產物為quinolinic acid，目前被認為具有活化N-methyl-D-aspartate (NMDA) 受體的能力。在過去的研究中，因為KMO下游產物會造成中樞神經過度興奮而導致神經毒性，故主要被認為與神經退化性疾病有關(例如: Alzhemer’s disease and Huntington’s disease)；然而在我們過去對犬乳腺腫瘤的研究中發現， KMO的表現量分別與腫瘤的惡性程度和病畜生存時間具有相關性，其中越惡性或級數越高的犬乳腺腫瘤其KMO表現量越高，且KMO表現量越高的病畜其存活時間則越短，但KMO在犬乳腺腫瘤細胞中的功能，目前仍不清楚，故本篇研究主要在探討KMO於犬乳腺腫瘤細胞中的功能。首先，我們確認實驗室現有的三株犬乳腺腫瘤細胞株，CMT-1, MPG及CF41.mg皆有表現KMO的mRNA及蛋白質，且在投與專一性的KMO抑制劑(Ro 61-8048)後，KMO蛋白質的表現並不會受到影響。其次，投與該KMO抑制劑，可以抑制犬乳腺腫瘤細胞的增生及移行能力，而在同時投與NMDA受體的致效劑NMDA後，能夠有效的反轉KMO抑制劑造成抑制腫瘤細胞增生能力，但卻不能逆轉被抑制的腫瘤細胞移行能力。另外，在腫瘤細胞訊息傳遞的分析上，我們發現投與KMO抑制劑抑制KMO功能時，降低了腫瘤細胞內ERK, AKT及STAT3等分子的活性，而同時投與NMDA則反轉了ERK及STAT3的活性抑制作用，但不影響AKT的活性抑制作用。顯示KMO可能是藉由ERK及STAT3細胞傳訊路徑影響犬乳腺腫瘤細胞增生能力，而細胞移行能力則可能是藉由AKT細胞傳訊路徑控制。 | zh_TW |
| dc.description.abstract | Kynurenine 3-monooxygenase (KMO), mainly found on the outer membrane of the mitochondria, is one of the enzymes in the kynurenine pathway. KMO catalyzes L-kynurenine to form 3-hydroxy kynurenine and further generates the downstream metabolite quinolinic acid which is an N-methyl-D-aspartate (NMDA) agonist. In the past, KMO was used to associate with neurodegenerative diseases because the downstream metabolites demonstrate excitotoxicity to CNS and act as factors in neurodegenerative diseases. In our previous study has shown that overexpression of KMO is related to tumor malignancy and survival time of cMGT-suffering dogs (in progress). In this study, we investigated the functional characterization of KMO in cMGT cells. First of all, KMO expression of cMGT cells (CMT-1, MPG and CF41.mg) was identified in mRNA and protein level and not changed by application of KMO inhibitor. Second, proliferation and migration of cMGT cells was reduced by KMO inhibitor, and the inhibition of proliferation was restored by NMDA. Third, downregulation of KMO function by KMO inhibitor decreased ERK, AKT and STAT3 activity of cMGTs, and the activity of ERK and STAT3 was restored by NMDA. Our data suggest that KMO involves in the proliferation and migration of cMGT cells. Furthermore, the inhibition of proliferation by KMO inhibitor is mainly through ERK and STAT3 signaling cascades and the signaling pathway may be activated through the NMDA receptor. The inhibition of migration by KMO inhibitor is mainly through AKT signaling cascades. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T00:04:47Z (GMT). No. of bitstreams: 1 ntu-102-R00629023-1.pdf: 2404056 bytes, checksum: 6bc8d5f93121cb6de58c5dd615679a34 (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | Contents
口試委員審定書 I 誌謝 II 中文摘要 IV Abstract V Contents VII Chapter 1. Background and Literature Review 1 1.1 Kynurenine 3-monooxygenase 1 1.2 Canine mammary gland tumor 3 1.2.1. Prevalence of cMGTs 3 1.2.2. Classification and staging of cMGTs 4 1.2.3. Therapy for cMGTs 5 1.2.4. Molecular carcinogenesis of cMGTs 7 1.3 Kynurenine pathway 10 1.3.1. The function of kynurenine pathway in central nerve system (CNS) 10 1.3.2. The function of kynurenine pathway in immune system 12 1.3.3. The relationship of kynurenine pathway and tumors 14 1.4 Conclusion 17 Chapter 2. Introduction 19 Chapter 3. Materials and methods 23 3.1 Cell lines and reagents 23 3.2 Reverse transcription polymerase chain reaction (RT-PCR) 23 3.3 Western blotting 25 3.4 Immunocytochemistry 27 3.5 Proliferation assay 28 3.6 Cell cycle 28 3.7 Migration assay 29 3.8 Statistical analysis 30 Chapter 4. Results 31 4.1 KMO expression of cMGT cells was identified in mRNA and protein level 31 4.2 KMO expression of cMGT cells was not changed by application of KMO inhibitor 31 4.3 KMO inhibitor inhibited the migration ability in CMT-1 and MPG 32 4.4 KMO inhibitor inhibited the cell proliferation in CMT-1 and MPG 33 4.5 The inhibition of cell proliferation by KMO inhibitor was restored by NMDA agonist in cMGT cells 34 4.5 The effect of KMO inhibitor in combined with NMDA on cell migration in cMGT cells 34 4.6 Inhibition of KMO function by KMO inhibitor decreased ERK, AKT and STAT3 activity of CMT-1 35 4.7 The phosphorylative reduction of ERK and STAT3 by KMO inhibitor was restored by NMDA agonist in CMT-1 36 Chapter 5. Discussion 37 Tables 44 Table 1. Histological classification of canine mammary gland tumors (adapted from the study of Misdorp et al. 1999) 44 Table 2. Staging system of canine mammary gland tumors 45 Table 3. Correlation between the enzymes, metabolites or target receptors in kynurenine pathway and tumor malignancy 46 Table 4. Sequence of primers used in RT-PCR 48 Figures 49 Figure 1. The kynurenine metabolites pathway and the target receptor 49 Figure 2.The mRNA and protein expression of KMO in cMGT cells 50 Figure 3. Effect of KMO expression on cMGT cells by application of KMO inhibitor 52 Figure 4. Effect of KMO inhibitor on the motility of cMGT cells 54 Figure 5. Effect of KMO inhibitor and IDO inhibitor on the cell viability of cMGT cells 55 Figure 6. Effect of KMO inhibitor on the cell cycle in CMT-1 56 Figure 7. NMDA receptor agonist (NMDA) reverses the antiproliferation effect of KMO inhibitor on cMGT cells 57 Figure 8. The effect of KMO inhibitor on ERK1/2, AKT and STAT3 signal pathways 58 Figure 9. The effect of KMO inhibitor in combination with NMDA on migration ability in cMGT cells 60 Figure 10. The effect of the combination of KMO inhibitor and NMDA on cell signal pathway in cMGT cells 61 Figure 11. Scheme of the mechanism KMO involved in cMGT cells 62 Reference 63 | |
| dc.language.iso | en | |
| dc.title | 探討KMO於犬乳腺腫瘤細胞的功能 | zh_TW |
| dc.title | Functional Characterization of Kynurenine 3-monooxygenase in Canine Mammary Gland Tumors (cMGT) Cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 郭村勇,施能耀,王愈善,林辰栖 | |
| dc.subject.keyword | Kynurenine 3-monooxygenase (KMO),kynurenine pathway,犬乳腺腫瘤,Ro 61-8048, | zh_TW |
| dc.subject.keyword | Kynurenine 3-monooxygenase (KMO),kynurenine pathway,canine mammary gland tumor (cMGT),KMO inhibitor (Ro 61-8048), | en |
| dc.relation.page | 76 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2013-08-14 | |
| 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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