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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 | 張逸良,施金元 | |
dc.contributor.author | Jheng-Cheng Huang | en |
dc.contributor.author | 黃正誠 | zh_TW |
dc.date.accessioned | 2021-06-15T16:25:52Z | - |
dc.date.available | 2020-09-25 | |
dc.date.copyright | 2015-09-25 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-08-14 | |
dc.identifier.citation | 1 衛生福利部。103年國人死因統計結果。行政院衛生福利部 <http://www.mohw.gov.tw/cht/Ministry/DM2_P.aspx?f_list_no=7&fod_list_no=5313&doc_no=49778> (2015).
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Nat Med 19, 1389-1400, doi:10.1038/nm.3388 http://www.nature.com/nm/journal/v19/n11/abs/nm.3388.html#supplementary-information (2013). 7 Sharma, S. V., Bell, D. W., Settleman, J. et al. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer 7, 169-181 (2007). 8 Gazdar, A. F. Activating and resistance mutations of EGFR in non-small-cell lung cancer: role in clinical response to EGFR tyrosine kinase inhibitors. Oncogene 28, S24-S31, doi:10.1038/onc.2009.198 (2009). 9 Sequist, L. V., Bell, D. W., Lynch, T. J. et al. Molecular Predictors of Response to Epidermal Growth Factor Receptor Antagonists in Non–Small-Cell Lung Cancer. Journal of Clinical Oncology 25, 587-595 (2007). 10 Gainor, J. F. & Shaw, A. T. Emerging Paradigms in the Development of Resistance to Tyrosine Kinase Inhibitors in Lung Cancer. Journal of Clinical Oncology 31, 3987-3996 (2013). 11 MacDonald, B. T., Tamai, K. & He, X. Wnt/β-catenin signaling: components, mechanisms, and diseases. Developmental cell 17, 9-26, doi:10.1016/j.devcel.2009.06.016 (2009). 12 Xi, Y. & Chen, Y. Wnt signaling pathway: Implications for therapy in lung cancer and bone metastasis. Cancer Letters 353, 8-16, doi:http://dx.doi.org/10.1016/j.canlet.2014.07.010 (2014). 13 Stewart, D. J. Wnt Signaling Pathway in Non–Small Cell Lung Cancer. Journal of the National Cancer Institute 106 (2014). 14 Takahashi, N., Maeda, K., Ishihara, A. et al. Regulatory mechanism of osteoclastogenesis by RANKL and Wnt signals. Front Biosci (Landmark Ed) 16, 21-30 (2011). 15 Prgomet, Z., Axelsson, L., Lindberg, P. et al. Migration and invasion of oral squamous carcinoma cells is promoted by WNT5A, a regulator of cancer progression. Journal of Oral Pathology & Medicine, n/a-n/a, doi:10.1111/jop.12292 (2014). 16 Shojima, K., Sato, A., Hanaki, H. et al. Wnt5a promotes cancer cell invasion and proliferation by receptor-mediated endocytosis-dependent and -independent mechanisms, respectively. Scientific Reports 5, 8042, doi:10.1038/srep08042 (2015). 17 Lin, L., Liu, Y., Zhao, W. et al. Wnt5A expression is associated with the tumor metastasis and clinical survival in cervical cancer. International Journal of Clinical and Experimental Pathology 7, 6072-6078 (2014). 18 Hung, T.-H., Hsu, S.-C., Cheng, C.-Y. et al. Wnt5A regulates ABCB1 expression in multidrug-resistant cancer cells through activation of the non-canonical PKA/β-catenin pathway. Oncotarget 5, 12273-12290 (2014). 19 Wei W, S. H., Li N, Li HY, Li X, Li Q, Shen XH. WNT5A modulates cell cycle progression and contributes to the chemoresistance in pancreatic cancer cells. Hepatobiliary Pancreat Dis Int 13, 529-538, doi:10.1016/s1499-3872(14)60277-0 (2014). 20 Bieche, I., Onody, P., Laurendeau, I. et al. Real-time reverse transcription-PCR assay for future management of ERBB2-based clinical applications. Clin Chem 45, 1148-1156 (1999). 21 Yuan, A. N. G., Yang, P.-C., Yu, C.-J. et al. Interleukin-8 Messenger Ribonucleic Acid Expression Correlates with Tumor Progression, Tumor Angiogenesis, Patient Survival, and Timing of Relapse in Non–Small-Cell Lung Cancer. American Journal of Respiratory and Critical Care Medicine 162, 1957-1963, doi:10.1164/ajrccm.162.5.2002108 (2000). 22 Yoon, S., Choi, M.-h., Chang, M. S. et al. Wnt5a-Dopamine D2 Receptor Interactions Regulate Dopamine Neuron Development via Extracellular Signal-regulated Kinase (ERK) Activation. Journal of Biological Chemistry 286, 15641-15651 (2011). 23 Jannesari-Ladani, F., Hossein, G., Monhasery, N. et al. Wnt5a influences viability, migration, adhesion, colony formation, E- and N-cadherin expression of human ovarian cancer cell line SKOV-3. Folia Biol (Praha) 60, 57-67 (2014). 24 Dissanayake, S. K., Wade, M., Johnson, C. E. et al. The Wnt5A/protein kinase C pathway mediates motility in melanoma cells via the inhibition of metastasis suppressors and initiation of an epithelial to mesenchymal transition. J Biol Chem 282, 17259-17271, doi:10.1074/jbc.M700075200 (2007). 25 Scheel, C., Eaton, E. N., Li, S. H. et al. Paracrine and autocrine signals induce and maintain mesenchymal and stem cell states in the breast. Cell 145, 926-940, doi:10.1016/j.cell.2011.04.029 (2011). 26 Gujral, T. S., Chan, M., Peshkin, L. et al. A noncanonical Frizzled2 pathway regulates epithelial-mesenchymal transition and metastasis. Cell 159, 844-856, doi:10.1016/j.cell.2014.10.032 (2014). 27 Wilson, C., Nicholes, K., Bustos, D. et al. Overcoming EMT-associated resistance to anti-cancer drugs via Src/FAK pathway inhibition. Oncotarget; Vol 5, No 17 (2014). | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/52748 | - |
dc.description.abstract | 肺癌是癌症死亡排行榜的首位。研究報告顯示表皮生長因子受體 (epidermal growth factor receptor, EGFR) 的突變在東方人的肺癌細胞有較高比例。EGFR是一個酪胺酸激酶受體 (receptor tyrosine kinase),與肺癌細胞的分化、增生、轉移及存活有關。目前國內的肺癌病患若是檢測出具有EGFR mutations (exon 19 deletion or/and L858R),將以表皮生長因子受體激酶抑制劑 (EGFR-TKI) 作為標靶治療的藥物。目前可以使用的EGFR-TKI包括有 Gefitinib (Iressa) 與 Erlotinib (Tarceva) 以及 Afatinib (Gilotrif)。然而EGFR突變的病患平均在標靶治療後的8到12個月就會出現抗藥性,因此仍須尋找EGFR-TKI產生抗藥性的相關基因。目前此實驗初步的研究結果顯示在gefitinib-resistant cell line (PC9/gef) and sensitive cell line (PC9) 透過cDNA microarray分析中找到與EGFR-TKI抗藥性相關的Wnt5a基因。Wnt5a是Wnt家族19個成員的其中之一,Wnt配體由高保守性的醣蛋白組成,能夠調節各種細胞功能,包括增殖、存活以及遷移。有研究報告指出Wnt5a與癌症病患的預後不佳有關係。根據本研究結果表示相較於EGFR-TKI-sensitive PC9 細胞株,Wnt5a在EGFR-TKI-resistant PC9/gef的細胞株表現有明顯的上升。當我們在PC9/gef細胞株抑制了Wnt5a的表現時將可以觀察到細胞對於EGFR-TKI的抗藥性、細胞生長、遷移以及入侵能力受到抑制。這可能暗示著Wnt5a在肺癌細胞的功能與EGFR-TKI抗藥性過程中扮演重要的角色。 | zh_TW |
dc.description.abstract | Lung cancer is the leading cause of cancer death. Some studies indicate that epidermal growth factor receptor (EGFR) mutation in Asian lung cancer cell of a substantial percentage. EGFR is a receptor tyrosine kinase that correlates with cell proliferation, growth, metastasis and survival in lung cancer. Currently, EGFR-TKI target therapy is specific for patients with exon 19 deletion or/and L858R mutations. EGFR-TKIs include the drugs Gefitinib (Iressa), Erlotinib (Tarceva) and Afatinib (Gilotrif). Gefitinib and Erlotinib are the first-generation reversible EGFR TKIs. Afatinib is the second-generation irreversible EGFR TKI. Despite of high response rate in these patients, the resistance to EGFR-TKIs occurs on average between 8-12 months. Therefore, we tried to find genes capable of conferring EGFR-TKI resistance. In our studies, we performed cDNA microarrays comparing gefitinib-resistant cell line (PC9/gef) and parental sensitive cell line (PC9) to identify the resistance-related genes, which may be involved in the EGFR-TKI resistance. The candidate gene Wnt5a (Wingless-type MMTV integration site family, member 5A) is a component of Wnt signaling, and we are interested in studying the role of Wnt5a in EGFR-TKI resistance. Wnt ligands consist of 19 highly conserved secreted glycoproteins that regulate various of cell functions including proliferation, survival, and migration. Although some studies indicate that Wnt5a correlates with poor overall survival in patients with lung cancer, the role of Wnt5a in relation to EGFR-TKIs resistance remains unclear. In our results, significant up-regulation of Wnt5a mRNA and protein in EGFR-TKI resistant PC9/gef as compared to its expression in EGFR-TKI sensitive PC9 cell line. We found that EGFR-TKI resistance decreased by the knockdown of Wnt5a in EGFR-TKI resistant cells. At the same time we also found that cell proliferation, migration and invasion ability were decreased by the knockdown of Wnt5a in EGFR-TKI resistant cells. These results implied that Wnt5a played a critical role in EGFR-TKI resistance and the functions of lung cancer. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T16:25:52Z (GMT). No. of bitstreams: 1 ntu-104-R02444002-1.pdf: 2003823 bytes, checksum: 1204d70772132d7c1b0ead047cf267cc (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 口試委員審定書……………………………………………………….………… #
中文摘要…………………………………………………………………….…… 1 Abstract………………………………………………………………….……….. 2 目錄………………………………………………………………………….…… 4 第一章、緒論 Introduction…………………………………………….……… 6 1.1 肺癌 (Lung cancer) ……………………………………….…….......... 6 1.2 表皮生長因子受體 (epidermal growth factor receptor, EGFR)….….. 6 1.3 酪胺酸激酶抑制劑 (tyrosine kinase inhibitors, TKI)……...…….…... 8 1.4 Wnt5a (Wingless-type MMTV integration site family, member 5A)…. 8 1.5 Wnt5a與癌症 (Wnt5a and Cancer)…………..………………………. 10 1.6 實驗目標………..…………………..…………………..………..……. 10 第二章、實驗材料及方法 Materials and methods…………………………… 11 2.1 細胞株與細胞培養 (Cell lines and culture conditions) ………….…... 11 2.2 細胞增生試驗 (Cell proliferation assay) …………...………...……… 11 2.3 基因靜默 (Gene silencing) ………………...…….…………………… 11 2.4 即時定量反轉錄聚合酶連鎖反應 (Real-time Quantitative reverse transcriptase RT-qPCR) ……………………………….………………. 12 2.5 西方墨點法 (Western blotting)……………………………………….. 12 2.6 細胞存活率分析 (Cell viability assays)……………………..……..…. 13 2.7 細胞遷移和侵入能力試驗 (Cell migration and invasion assay) ….…. 14 2.8 組織來源 (Tissue Procurement) …………………………………...….. 14 2.9 免疫組織化學染色法 (Immunohistochemistry) ……….…………….. 15 2.10 統計分析 (Statistical analyses)……...………………………………… 17 第三章、結果 Results…………………………………………………………… 18 3.1 利用cDNA microarray預測EGFR-TKI抗藥性的相關基因………... 18 3.2 Wnt5a在其他抗藥性的肺癌細胞株的表現量差異………….…...…... 18 3.3 抑制Wnt5a表現與EGFR-TKI抗藥性的關係…………….…….…… 19 3.3.1 利用siRNA在PC9/gef抗藥性細胞株抑制Wnt5a表現的效率.. 19 3.3.2 抑制Wnt5a的表現增加細胞對於Erlotinib或Afatinib的敏感性 19 3.4 分析Wnt5a在肺癌細胞的功能……….………………………….…… 20 3.4.1 抑制Wnt5a降低細胞遷移能力 (migration ability)……………... 20 3.4.2 抑制Wnt5a降低細胞侵入能力 (invasion ability)…………..…... 21 3.4.3 抑制Wnt5a減少細胞增生能力 (proliferation ability)…..….…... 21 3.5 肺癌病患檢體Wnt5a的表現與EGFR TKIs之相關性……….……… 22 第四章、討論 Discussions………………………..…………..……….………… 23 第五章、實驗圖表 Figures and Tables…………………………….…………… 26 第六章、參考文獻 References…………………………………………………. 37 | |
dc.language.iso | zh-TW | |
dc.title | 探討Wnt5a在肺癌細胞的功能與EGFR-TKI抗藥性的關聯 | zh_TW |
dc.title | Investigation of the Wnt5a function in lung cancer and the correlation between Wnt5a and TKI-resistance | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 蔡孟峰 | |
dc.subject.keyword | 肺癌,EGFR-TKI,抗藥性,Wnt5a, | zh_TW |
dc.subject.keyword | Lung cancer,EGFR-TKI,Resistance,Wnt5a, | en |
dc.relation.page | 41 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2015-08-14 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 病理學研究所 | zh_TW |
顯示於系所單位: | 病理學科所 |
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