https://scholars.lib.ntu.edu.tw/handle/123456789/164209
標題: | 犬淋巴瘤細胞表現型鑑定以及治療藥物之評估 Identification of Phenotype and Evaluation of Therapeutic Drugs on Canine Lymphoma Cells |
作者: | 吳瑞紘 Wu, Rui-Hong |
關鍵字: | 犬淋巴瘤;表型分析;流式細胞技術;烷基化藥物;Kynurenine 3-monooxygenase (KMO);dovitinib | 公開日期: | 2014 | 摘要: | 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。 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. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/264485 | Rights: | 論文公開時間:2016/08/08 論文使用權限:同意有償授權(權利金給回饋本人) |
顯示於: | 獸醫學系 |
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ntu-103-R01629004-1.pdf | 23.32 kB | Adobe PDF | 檢視/開啟 |
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