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  1. NTU Theses and Dissertations Repository
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請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17278
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor林辰栖
dc.contributor.authorYu-Ya Huangen
dc.contributor.author黃郁雅zh_TW
dc.date.accessioned2021-06-08T00:04:34Z-
dc.date.copyright2013-08-28
dc.date.issued2013
dc.date.submitted2013-08-14
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17278-
dc.description.abstractX814是常見的中草藥,在近年來許多研究發現其具有多重作用,包含免疫調節、抗腫瘤、抗微生物、抗血管新生與刺激免疫反應等。而目前並沒有任何文獻探討X814在犬隻腫瘤上的影響。犬乳腺腫瘤為母犬中最為好發的腫瘤。七歲以上患有腫瘤的母狗中,有52%以上的機率為乳腺腫瘤,而其中又有50%的腫瘤為惡性。手術為犬乳腺腫瘤的主要治療方法,但是術後依然有50%的機率會發生復發或轉移。因此,發展新的、具抗犬乳腺腫瘤的藥物相當的重要。本研究旨在探討X814的粗萃物,以酒精與正己烷溶劑製備後,評估其在犬乳腺腫瘤上的抗腫瘤效果。結果顯示該藥的酒精萃取物(X.E.E.)與正己烷萃取物(X.H.E.)能引起犬乳腺腫瘤細胞發生壞死性死亡、自體吞噬、與細胞周期停滯;也會抑制細胞聚落形成、移行與增生的能力。在蛋白質表現方面,X.E.E.與X.H.E.會增加犬乳腺腫瘤細胞的LC3Ⅱ蛋白的表現,而在許多文獻皆指出LC3Ⅱ的表現量與細胞自體吞噬作用具有正相關。另外,細胞質中的p53與p-AKT的表現皆有下降,可能與細胞壞死性死亡、自體吞噬、細胞周期停滯、細胞移行能力下降有關。由於細胞自體吞噬的作用在腫瘤上扮演著一體兩面的腳色,我們進一步探討X.E.E.與X.H.E.所引起細胞自體吞噬的影響。研究結果顯示以自體吞噬抑制劑bafilomycin A1或chloroquine與X.E.E.或X.H.E.共同作用後,能更加強腫瘤細胞的死亡效果。前述結果顯示X814萃取物X.E.E.與X.H.E.具有治療犬乳腺腫瘤的潛力。zh_TW
dc.description.abstractX814 is an herbal plant shown to possess a number of beneficial anti-cancer and immunomodulatory properties in recent reports. Though potential anti-microbial, anti-angiogenesis, and immune-stimulation activities have been demonstrated, the effects of X814 on canine tumors have not been well elucidated. Canine mammary gland tumors (cMGT) are the most frequently diagnosed neoplasms in female dogs. Roughly 52% of canine mammary tumors are diagnosed in female dogs older than seven years. About 50% of the cases are diagnosed as malignant mammary carcinoma. Although surgical excision is now viewed as the most effective modality for treatment of cMGT, 50% of them relapsed after surgery, and the two-year survival rate of malignant cases were less than 50%. Therefore, developing potential anti-cMGT therapies becomes much more important. The crude extracts of X814 were firstly prepared in ethanol and hexane solvents to evaluate their anti-proliferation effects on cMGT cell lines. The results showed both the EtOH extract (X.E.E.) and hexane extract (X.H.E.) of X814 could inhibit cell proliferation, colony formation, and cell migration in cMGT cells. The reduction of tumor cell growth was resulted from necrosis, autophagy, and cell cycle arrest. Moreover, cMGT cells treated with X.E.E. and X.H.E. showed increased expression of autophagy-related proteins, LC3Ⅱ, and decreased cytoplasmic p53 and p-AKT in a dose-dependent manner. Autophagy seemed to play a protective role in X.E.E and X.H.E.-treated cMGT cells since using autophagy inhibitor, bafilomycin A1 or chloroquine, enhanced the cell death. Collectively, our results indicated that X814 has potential anticancer effects on cMGT.en
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dc.description.tableofcontents審查書 I
誌謝 II
中文摘要 III
Abstract IV
Chapter 1 Background 1
1.1 Cancer review 1
1.1.1 Cancer definition 1
1.1.2 The mechanism of cancer arise 1
1.1.3 Cancer therapy 3
1.2 Canine mammary gland tumors (cMGT) 5
1.2.1 The influence of incidence, age, breed and sexual hormone influence on canine mammary gland tumors (cMGT) 5
1.2.2 Presentation and clinical signs of canine mammary gland tumors (cMGT) 6
1.2.3 Histological classification of canine mammary gland tumors (cMGT) 7
1.2.4 Prognosis of canine mammary gland tumors (cMGT) 9
1.2.5 Treatment of canine mammary gland tumors (cMGT) 10
1.3 Traditional toxic Chinese medicines (TCM) review 11
1.3.1 Anticancer drugs from traditional toxic Chinese medicines (TCM) 11
1.3.2 Mechanisms of cancer therapy using traditional toxic Chinese medicines (TCM) 12
1.3.3 Cancer therapy by inducing cell death 14
1.3.3.1 Cell necrosis 14
1.3.3.2 Cell apoptosis 15
1.3.3.3 Cell autophagy 17
1.3.4 The function of p53, AKT and LC3 on cancer research 18
1.4 Euphorbiaceae family review 19
1.4.1 Hexane fraction of Euphorbiaceae family on antitumor activity 19
1.4.2 Biological functions of X814 20
Chapter 2 Strategy 22
Chapter 3 Materials and methods 23
3.1 Chemicals and reagents 23
3.2 Cell culture 23
3.3 Cell proliferation assay 24
3.4 Assessment of viable cell number 25
3.5 Migration assay 25
3.6 Soft agar assay for colony formation 26
3.7 Cell-mediated cytotoxicity assay 27
3.8 Annexin V assay 28
3.9 Sub G1 analysis 29
3.10 Cell staining with acridine orange 30
3.11 Preparation of conditioned media and western blot analysis 31
3.13 Statistical analysis 32
Chapter 4 Results 33
4.1 X.E.E. and X.H.E. inhibited cell proliferation of cMGT cells. 33
4.2 X.E.E and X.H.E inhibited cell proliferation by inducing cell death of cMGT cells 33
4.3 X.E.E. and X.H.E. reduced colony formation of cMGT cells 35
4.4 X.E.E. and X.H.E. inhibited cMGT cells migration 36
4.6 X.E.E. and X.H.E. induced no apoptosis on cMGT cells. 36
4.8 Autophagy was involved in X.E.E and X.H.E-treated cMGT cells 37
4.9 Blocking autophagy enhanced X.E.E. and X.H.E-mediated celll death 38
4.10 X.E.E. and X.H.E. increased LC3Ⅱ and decreased cytoplasmic p53 and p-AKT expression on cMGT cells. 39
Chapter 5 Discussion 40
Figures and tables 48
Reference 67
dc.language.isoen
dc.title探討X814在犬乳腺腫瘤上的抗癌效果zh_TW
dc.titleInvestigate the anti-tumor effect of X814 on canine mammary gland tumorsen
dc.typeThesis
dc.date.schoolyear101-2
dc.description.degree碩士
dc.contributor.oralexamcommittee廖泰慶,郭村勇,廖光文,王愈善
dc.subject.keyword犬乳腺腫瘤,自體吞噬,p53,p-AKT,zh_TW
dc.subject.keywordCanine mammary gland tumors (cMGT),autophagy,p53,p-AKT,en
dc.relation.page78
dc.rights.note未授權
dc.date.accepted2013-08-14
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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