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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李明亭(Ming-Ting Lee) | |
dc.contributor.author | Pei-Hsun Tsai | en |
dc.contributor.author | 蔡沛勳 | zh_TW |
dc.date.accessioned | 2021-06-16T10:18:26Z | - |
dc.date.available | 2018-08-28 | |
dc.date.copyright | 2013-08-28 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-16 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60447 | - |
dc.description.abstract | 癌症是已開發國家最主要的死亡原因之一,而癌症轉移則是癌症病患死亡的主因。當癌細胞開始轉移時,他們會形成侵襲性偽足(Invadopodia)、釋放基質金屬蛋白酶(MMP),接著穿透細胞外基質,脫離原位癌。近來有許多的文獻指出癌症幹細胞(CSC),因為其自我新生的能力、能分化成其他子代癌細胞,並可高度抵抗標準癌症治療,因此與腫瘤的復發和轉移有相當大的關聯。類黃鹼素(flavonoid)已被證實具有抗氧化、抗發炎、抗癌、抗癌轉移以及抗癌症幹細胞的功能。本實驗室先前挑選出兩種具潛力的類黃鹼素:木犀草素(Luteolin)與槲皮素(Quercetin),可抑制癌細胞的入侵、移動、MMP-9釋放並逆轉皮質-間葉細胞型態轉變(epithelial-mesenchymal transisiton, EMT)。
在本研究,我們將Du145攝護腺癌細胞(Du145-P),利用Boyden chamber進行篩選,經過連續三次穿膜篩選出Du145第三代細胞(Du145-III)。Du145-III展現了高度的腫瘤血管生成擬態、自我新生能力和癌症幹細胞標記蛋白。此外,Du145-III亦釋放高量的MMP-9,增強其移動和入侵的能力。在此,我們證實了木犀草素與槲皮素,可經由調控JNK訊息傳遞路徑,進而抑制Du145-III的MMP-9活性、移動、入侵、腫瘤血管生成擬態、自我新生能力,並降低癌症幹細胞標記蛋白的表現。 在另一主題的研究,我們利用相同的方式篩選出較高入侵能力的A431第三代細胞(A431-III)。A431-III擁有高度入侵和移動能力,並表現較多量的MMP-9和具有EMT的細胞型態。我們藉由A431-III和A431-P兩株細胞的比較,發現A431-III比起母代細胞形成較多的侵襲性偽足,並伴隨較高程度的細胞外基質分解。我們同時也觀察到cortactin和Src磷酸化程度的增強,在A431-III中調控著侵襲性偽足的形成。針對A431-III使用木犀草素與槲皮素,可透過降低cortactin 和Src的磷酸化,進而抑制侵襲性偽足的形成和釋放MMP-9的能力。 總結而言,我們建立了具高入侵能力的Du145和A431細胞模式系統,在探討癌症轉移中,可作為研究癌症幹細胞和侵襲性偽足的有用工具。同時我們亦證實了木犀草素與槲皮素,是針對癌症轉移與癌症幹細胞的有效藥物。 | zh_TW |
dc.description.abstract | Cancer is the leading cause of death in developed country. Metastasis is a major cause of mortality in cancer patients. When cancer cells start to invade, they could form invadopodia structure, secret matrix metalloproteinases (MMPs) and then penetrate across extracellular matrix to depart from primary tumor. Recent studies indicate that cancer stem cells (CSCs) are involved in tumor recurrence and metastasis, owing to their self-renewal capability, differentiation to cancer progeny cells, and higher resistance to canonical therapy.
Flavonoids have been documented to exhibit anti-oxidative, anti-inflammatory, anti-carcinogenic and anti-metastatic effects as well as anti-CSCs. Previously, we found that two potent flavonoids, luteolin (Lu) and quercetin (Qu), could inhibit invasion, migration, MMP-9 secretion and induce reversal of epithelial-mesenchymal transition (EMT). In this study, we have established a prostate cancer sub-line Du145-III, which was derived from parental Du145 (Du145-P) cells by three successive passages through Boyden chamber. Du145-III exhibits a higher level of vasculogenic mimicry (VM), self-renewal capability and CSC markers. In addition, Du145-III also secretes relatively greater quantity of MMP-9 leading to enhanced migratory and invasive abilities. Here, we demonstrate that Lu and Qu, targeting JNK signaling pathway, could inhibit MMP-9 activity, migratory, invasion, VM, self-renewal capability, and suppress expression of CSC markers. In a separate study, we isolated a highly invasive A431-III sub-line using the same method described above. The A431-III possesses higher invasive and migratory abilities, elevated levels of MMP-9 and an epithelial-mesenchymal transition (EMT) phenotype. Using this A431-III sub-line, we discovered that A431-III cells had an increased potential to form invadopodia accompanied by higher ECM degradation ability compared with A431-P. We also observed enhanced phosphorylation levels of cortactin and Src which regulate invadopodia formation in A431-III cells. Treatment of A431-III cells with Lu and Qu, caused attenuation the phosphorylation of cortactin and Src, then inhibiting invadopodia formation and MMP-9 secretion. In summary, we established highly invasive Du145 and A431 model system serve as a useful tool to investigate the role of both CSC and invadopodia formation in cancer metastasis. We also provide evidence indicating that Lu and Qu are powerful agents to target cancer invasion and CSC. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T10:18:26Z (GMT). No. of bitstreams: 1 ntu-102-D96b46008-1.pdf: 4372243 bytes, checksum: bd6d9314b3a5ce928728e8ce11bef514 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 中文摘要 i
Abstract iii Content v List of figures vii PART I 1 Introduction 2 Matrix metalloproteinases 4 Vasculogenic mimicry 5 Cancer stem cell 7 Cancer stem cells and metastasis 9 Identification of cancer stem cells 9 Signaling pathway involved in cancer stem cells 11 Flavonoids 13 Purpose and importance 15 Materials and Methods 17 Materials 17 Cell culture 18 In vitro invasion assays and isolation of highly invasive Du145 tumor cells 18 Preparation of cell lysates 19 Detection and characterization of MMPs by gelatin zymography 19 In vitro wound-healing migration assay 20 In vitro capillary formation assay 21 Co-culture system of Du145-III and HUVEC in capillary formation assay 21 Western blot 22 Reverse transcriptase-polymerase chain reaction (RT-PCR) 22 Transfection of small interference RNA (siRNA) 23 Spheroid formation assay 23 Statistical analysis 24 Results 25 Du145-III, a highly invasive cancer sub-line, is isolated from Du145 prostate cancer cell 25 Du145-III cells exhibit vasculogenic mimicry and integrate into HUVEC network 26 Du145-III cells possess higher self-renewal capability and expression of CSC markers than Du145-P cells 27 MMP-9 is involved in the self-renewal and vasculogenic mimicry in Du145-III cells 29 JNK signaling pathway is associated with maintenance of stemess and metastatic potential in Du145-III cells 30 Luteolin and Quercetin inhibit the vasculogenic mimicry, metastatic potential and stemness via JNK activity in Du145-III cells 32 Discussion 52 PART II 59 Introduction 60 Invasion of cancer cell 60 Matrix metalloproteinase 62 Invadopodia 64 Cortactin 66 Flavonoids 68 Purpose and importance 71 Materials and Methods 73 Materials 73 Cell culture 74 Preparation of cell lysate 74 Transfection of small interference RNA 75 Western blot 76 Gene expression microarray analysis 76 Quantitative real time PCR 77 Gelatin zymography 77 Immunofluorescence 78 Matrix degradation assay 79 Statistical analysis 80 Results 81 A431-III cells form more invadopodia and degrade the ECM effectively 81 Cortactin phosphorylation promotes invadopodia formation and matrix degradation. 83 MMP activity is required for invadopodia formation and matrix degradation 85 MMP-9 is the major MMP for invadopodia in A431-III cells 86 Luteolin and quercetin impair invadopodia formation and function in A431-III cells 87 Luteolin and quercetin inhibit Src kinase activity and suppress MMPs secretion 89 Discussion 101 Appendix 110 References 121 | |
dc.language.iso | en | |
dc.title | 木犀草素與槲皮素對高入侵性Du145和A431子代癌細胞致癌能力之影響
第一部份: 幹細胞特性與腫瘤血管生成擬態之抑制 第二部份: 基質金屬蛋白酶分泌及侵襲性偽足形成能力之抑制 | zh_TW |
dc.title | Anti-cancer effect of two flavonoids luteolin and quercetin on highly invasive Du145-III and A431-III cancer cell sublines
PART I : Suppression of stemness and vasculogenic mimicry PART II: Suppression of MMP-9 secretion and invadopodia formation | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 張震東(Geen-Dong Chang),黃銓珍(Chang-Jen Huang),陳宏文(Hungwen Chen),柯逢春(Fon-Chun Ke),黃娟娟(Jiuan-Jiuan Hwang) | |
dc.subject.keyword | 癌症轉移,癌症幹細胞,腫瘤血管生成擬態,侵襲性偽足,基質金屬蛋白酶,類黃鹼素, | zh_TW |
dc.subject.keyword | cancer metastasis,cancer stem cell,MMP-9,vasculogenic mimicry,invadopodia,flavonoids, | en |
dc.relation.page | 137 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-08-17 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科學研究所 | zh_TW |
顯示於系所單位: | 生化科學研究所 |
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