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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 黃敏銓(Min-Chuan Huang) | |
dc.contributor.author | Syue-Ting Chen | en |
dc.contributor.author | 陳學亭 | zh_TW |
dc.date.accessioned | 2021-06-17T05:00:08Z | - |
dc.date.available | 2023-08-01 | |
dc.date.copyright | 2018-08-01 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-07-25 | |
dc.identifier.citation | 1. Siegel RL, Miller KD, Jemal A: Cancer Statistics, 2017. CA Cancer J Clin 2017, 67:7-30.
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Watanabe S, Kishimoto T, Yokosuka O: Hepatocyte growth factor inhibits anoikis of pancreatic carcinoma cells through phosphatidylinositol 3-kinase pathway. Pancreas 2011, 40:608-614. 38. Rizwani W, Allen AE, Trevino JG: Hepatocyte Growth Factor from a Clinical Perspective: A Pancreatic Cancer Challenge. Cancers (Basel) 2015, 7:1785-1805. 39. Swartz MJ, Batra SK, Varshney GC, Hollingsworth MA, Yeo CJ, Cameron JL, Wilentz RE, Hruban RH, Argani P: MUC4 expression increases progressively in pancreatic intraepithelial neoplasia. Am J Clin Pathol 2002, 117:791-796. 40. Yamasaki H, Ikeda S, Okajima M, Miura Y, Asahara T, Kohno N, Shimamoto F: Expression and localization of MUC1, MUC2, MUC5AC and small intestinal mucin antigen in pancreatic tumors. Int J Oncol 2004, 24:107-113. 41. Moniaux N, Junker WM, Singh AP, Jones AM, Batra SK: Characterization of human mucin MUC17. Complete coding sequence and organization. J Biol Chem 2006, 281:23676-23685. 42. Takikita M, Altekruse S, Lynch CF, Goodman MT, Hernandez BY, Green M, Cozen W, Cockburn M, Sibug Saber M, Topor M, et al: Associations between selected biomarkers and prognosis in a population-based pancreatic cancer tissue microarray. Cancer Res 2009, 69:2950-2955. 43. Haridas D, Chakraborty S, Ponnusamy MP, Lakshmanan I, Rachagani S, Cruz E, Kumar S, Das S, Lele SM, Anderson JM, et al: Pathobiological implications of MUC16 expression in pancreatic cancer. PLoS One 2011, 6:e26839. 44. Chauhan SC, Ebeling MC, Maher DM, Koch MD, Watanabe A, Aburatani H, Lio Y, Jaggi M: MUC13 mucin augments pancreatic tumorigenesis. Mol Cancer Ther 2012, 11:24-33. 45. Goydos JS, Elder E, Whiteside TL, Finn OJ, Lotze MT: A phase I trial of a synthetic mucin peptide vaccine. Induction of specific immune reactivity in patients with adenocarcinoma. J Surg Res 1996, 63:298-304. 46. 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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71235 | - |
dc.description.abstract | 胰臟癌在十大癌症死因中排名第四位,大部份為胰腺癌。胰腺癌通常由胞外基質及星狀細胞共同形成的緻密纖維化基質所包覆,而由此纖維化基質所營造出的低養分,低氧及酸性的微環境促進了胰腺癌細胞的惡性行為。黏液蛋白(Mucins)為高度醣化之蛋白質,在惡性腫瘤的發病機轉中扮演重要角色。據研究顯示,各型的黏蛋白在胰腺癌中陸續被發現表現量異常,然而黏蛋白第二十型(MUC20) 在胰腺癌中扮演的角色仍未知。
本篇研究結果如下:胰腺癌組織的免疫染色結果與臨床特徵之相關性統計顯示,MUC20表現量高與低病人存活率及術後局部復發率呈正相關。血清剝奪、低氧及酸性環境皆能誘發胰腺癌細胞中MUC20的表現量。以小分子干擾核糖核酸抑制胰腺癌細胞株HPAC及HPAF-II 的MUC20表現量,則由星狀細胞所誘發的HPAC及HPAF-II爬行及侵襲能力也隨之減弱。在腹腔、皮下及胰臟注射胰腺癌細胞的動物模式中,降低胰腺癌細胞MUC20的表現量可減緩其在免疫缺陷鼠體內的生長。磷酸化-受體酪氨酸激酶陣列 (Phospho-Receptor Tyrosine Kinase Array, p-RTK array) 和西方墨點法 (Western Blot) 的分析結果顯示,將胰腺癌細胞中MUC20的表現量降低可同時降低由肝細胞生長因子(Hepatocyte Growth Factor, HGF)所誘發的肝細胞生長因子受體之磷酸化 (Phospho-Hepatocyte Growth Factor Receptor, p-MET)。由HGF所誘發的胰腺癌細胞惡性表徵,可藉由降低其細胞內MUC20的表現量而將之抑制。免疫共沉澱 (co-immunoprecipitation, co-IP) 的實驗結果顯示,MUC20和MET間有實質上的交互作用。 總結:胰腺癌細胞中MUC20表現量降低而產生其惡性表徵減弱的現象,至少部分是由於HGF/MET訊息傳遞路徑受到抑制而導致,同時也顯現MUC20極有潛力成為治療性標靶。 | zh_TW |
dc.description.abstract | Mucins are heavily glycosylated proteins that play critical roles in the pathogenesis of tumour malignancies. Pancreatic ductal adenocarcinoma (PDAC) is characterised by the aberrant expression of mucins. However, the role of mucin (MUC) 20 in PDAC remains unclear. PDAC is usually surrounded by a dense fibrotic stroma consisting of an extracellular matrix and pancreatic stellate cells (PSCs). The stroma creates a nutrient-deprived, hypoxic and acidic microenvironment and promotes the malignant behaviours of PDAC cells. In this study, immunohistochemical staining demonstrated that high MUC20 expression correlated with poor progression-free survival and high local recurrence rate of PDAC patients (n = 61). The expression of MUC20 was induced by serum deprivation, hypoxia, and acidic pH in PDAC cells. MUC20 knockdown with siRNA decreased cell viability as well as migration and invasion induced by PSCs in HPAC and HPAF-II cells. In intraperitoneal, subcutaneous, and orthotopic injection models, MUC20 knockdown decreased tumour growth in immunodeficient mice. Phospho-RTK array and western blot analysis indicated that MUC20 knockdown decreased HGF-mediated phosphorylation of MET in PDAC cells. Moreover, HGF-induced malignant phenotypes could be suppressed by MUC20 knockdown. Co-immunoprecipitation revealed the physical association of MUC20 and MET. These findings suggest that MUC20 knockdown suppresses the malignant phenotypes of PDAC cells at least partially through the inhibition of the HGF/MET pathway and that MUC20 could act as a potential therapeutic target. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T05:00:08Z (GMT). No. of bitstreams: 1 ntu-107-F01446009-1.pdf: 3133800 bytes, checksum: f56387f11fe59a8cba204fdddc84a5b6 (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | Table of contents
口試委員審定書 1 序言 2 中文摘要 3 Abstract 4 Chapter 1. Introduction 10 1.1. Pancreatic cancer 10 1.1A. Current status 10 1.1B. Precursors of PDACs 11 1.1C. Biomarker for early Detection 12 1.1D. Treatments 12 1.2. Microenvironment of PDAC 13 1.3. Receptor tyrosine kinases (RTKs) 14 1.3A. Background 14 1.3B. MET in PDAC 15 1.4. Mucins 16 Chapter 2. Materials and Methods 17 2.1. Immunohistochemistry 17 2.2. Cell lines and cell culture 17 2.3. cDNA synthesis and real time RT-PCR 18 2.4. Transfection and plasmid construction 19 2.5. Antibodies and reagents 20 2.6. MTT assay 21 2.7. Transwell migration and Matrigel invasion assays 21 2.8. In vivo xenograft tumour growth model 22 2.9. Phospho-receptor tyrosine kinase array assay 22 2.10. Immunoprecipitation 23 2.11. Statistical analysis 23 Chapter 3. Results 24 3.1. MUC20 is overexpressed in PDAC and MUC20 high expression correlates with poor survival and local recurrence 24 3.2. MUC20 knockdown inhibits 10% foetal bovine serum (FBS)-induced pancreatic cancer cell viability 25 3.3. MUC20 knockdown suppresses PDAC tumour growth in immunodeficient mice 26 3.4. MUC20 is up-regulated in the serum-deprived, hypoxic, and acidic microenvironment 27 3.5. MUC20 knockdown inhibits migration and invasion of PDAC cells co-cultured with PSCs 28 3.6. MUC20 enhances hepatocyte growth factor (HGF)/MET signalling in PDAC cells 29 3.7. Physical interactions of MUC20 and MET occur in PDAC cells 30 Chapter 4. Discussion 32 4.1. The role of MUC1 and MUC20 in PDAC 32 4.2. The role of HGF/MET signaling in interactions of PSC and PDAC 33 4.3. Modulation of the MUC20/HGF/MET signaling pathway under PDAC microenvironment 34 4.4. Interactions between MUC20 and MET 35 4.5. Effects of MUC20 overexpression on PDAC cells 35 Chapter 5. Conclusion 37 Chapter 6. References 38 Table of figures and legends Figure 1. MUC20 mRNA is overexpressed in PDAC and MUC20 high expression correlates with poor survival. 46 Figure 2. MUC20 expression in the primary PDAC tissue. 47 Figure 3. MUC20 protein is overexpressed in PDAC and MUC20 high expression correlates with poor progression free survival. 48 Figure 4. MUC20 expression in human normal and PDAC cell lines. 50 Figure 5. Effects of MUC20 knockdown on PDAC cells in vitro. 51 Figure 6. MUC20 knockdown suppresses PDAC tumour growth in immunodeficient mice. 52 Figure 7. MUC20 is up-regulated in serum-deprived, hypoxic and acidic microenvironment. 54 Figure 8. MUC20 is induced by serum deprivation. 55 Figure 9. The JNK signalling pathway is involved in MUC20 overexpression induced by serum deprivation in HPAC and HPAF-II cells. 56 Figure 10. MUC20 knockdown inhibits migration and invasion of PDAC cells co-cultured with PSCs. 57 Figure 11. MUC20 knockdown does not significantly affect 1% FBS-triggered migration and invasion in HPAC and HPAF-II cells. 59 Figure 12. MUC20 knockdown inhibits HGF/MET signalling in PDAC cells. 60 Figure 13. HGF mRNA is up-regulated by serum deprivation in PSCs. 61 Figure 15. MUC20 overexpression increases p-MET and cell migration triggered by HGF in HPAF-II cells. 63 Figure 16. MUC20 knockdown inhibits HGF-triggered cellular phenotypes in PDAC cells. 64 Figure 17. AKT overexpression increased HGF-triggered viability, migration, and invasion in stable MUC20 knockdown cells. 65 Figure 18. Physical interactions of MUC20 and MET occur in PDAC cells. 66 Figure 19. MUC20 levels in MUC20 knockdown cells are also lower than control cells under serum deprivation conditions. 67 Figure 20. MET expression in normal cell lines and PDAC cell lines. 68 Figure 21. MUC20 overexpression does not significantly affect PSC-triggered migration in MIA PaCa-2 cell. 69 Figure 22. Mucins expression in normal and PDAC cells analysed by real-time RT-PCR. 70 Figure 23. MET is up-regulated in serum-deprived, and acidic microenvironment. 71 Figure 24. Schematic diagram depicting how MUC20 mediates its biological effects through MET in PDAC cells. 72 Table of tables Table 1: Association between MUC20 expression with clinicopathologic characteristics of IPMN. 73 Table 2. Association between MUC20 expression with clinicopathologic characteristics of PDAC 74 Table 3. Primer sequence of Mucins for real-time RT-PCR. 75 | |
dc.language.iso | en | |
dc.title | 黏蛋白第二十型在胰臟癌中扮演的角色 | zh_TW |
dc.title | The role of MUC20 in pancreatic cancer | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 田郁文(Yu-Wen Tien) | |
dc.contributor.oralexamcommittee | 賴逸儒(I-Rue Lai),李明學(Ming-Shyue Lee),王淑慧(Shu-Huei Wang) | |
dc.subject.keyword | 胰腺癌,胰臟星狀細胞,黏液蛋白第二十型 (MUC20), | zh_TW |
dc.subject.keyword | Pancreatic ductal adenocarcinoma (PDAC),Pancreatic stellate cell (PSC),MUC20, | en |
dc.relation.page | 75 | |
dc.identifier.doi | 10.6342/NTU201801830 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-07-26 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 解剖學暨細胞生物學研究所 | zh_TW |
顯示於系所單位: | 解剖學暨細胞生物學科所 |
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