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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳惠文(Huei-Wen Chen) | |
dc.contributor.author | Li-Chieh Yu | en |
dc.contributor.author | 游力潔 | zh_TW |
dc.date.accessioned | 2021-06-08T00:14:31Z | - |
dc.date.copyright | 2013-09-24 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-07-31 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17462 | - |
dc.description.abstract | 肺癌為全球十大死因惡性腫瘤之一,在台灣發生率為所有惡性腫瘤中的第三名,死亡率則為為第一名,為威脅人民健康的重大疾病之一。肺癌可分為兩大類型:小細胞肺癌(small cell lung cancer)及非小細胞肺癌(non-small cell lung cancer, NSCLC),以非小細胞肺癌為大宗,約佔了85%的肺癌病患,其中又以肺腺癌(lung adenocarcinoma)佔NSCLC多數。造成肺腺癌的原因十分的複雜,環境汙染物、致癌基因的突變、遺傳等因素皆會影響肺腺癌的進程。
本篇研究的第一部分為探討環境汙染物-DEHP對於肺腺癌的影響,DEHP被發現與許多癌症的進程有關,但過去鮮少有探討塑化劑與肺腺癌關係的文獻。根據實驗結果,我們發現長期暴露DEHP會促使細胞轉移能力上升,在進一步利cDNA微陣列分析後發現DEHP會使調控細胞轉移、增生及黏附的基因表現量上升,其中一重要基因AGR2在過去的文獻研究中顯示與癌症進程有關,綜合以上的實驗結果,我們發現DEHP會促使AGR2表現量上升而使細胞轉移能力增強並提供DEHP影響肺癌的進程一分子機轉。 第二部分以代謝體學探討上皮生長因子(Epidermal growth factor receptor, EGFR)的突變對於肺腺癌細胞代謝之影響,並從中找出具代表此疾病的生物標記。L858R點突變為常見的EGFR突變形式。因此,針對EGFR所開發的藥物-酪胺酸激酶抑制劑(tyrosine kinase inhibitor, TKI)成為近年來治療肺腺癌的主流。然而,臨床上發現服用TKI後,病人會產生T790M的點突變會使得對EGFR-TKI治療產生抗藥性。因此,找出一生物標記作為診斷及用藥評估為一重要課題,我們利用代謝體學來分析此兩種EGFR突變小鼠肺癌組織中代謝途徑的差異,實驗結果顯示有二十個代謝物在EGFR突變小鼠肺臟中具顯著差異,且主要分布於糖解、檸檬酸循環、五碳糖循環、脂質代謝以及甲硫氨酸-回收途徑之中。我們建構出帶有EGFR突變的肺癌組織中其代謝體圖譜,並從中找出與細胞生長及癌症生成相關的代謝物如: spermine、hypoxanthine等。未來將會進一步分析病人血清來找出可以作為生物標記之代謝物。 根據本篇論文的研究顯示長期暴露DEHP會影響肺癌之進程,並利用代謝體學建立帶有EGFR突變的肺癌之代謝圖譜。 | zh_TW |
dc.description.abstract | Lung cancer is the leading cause of cancer mortality in worldwide. According to the Taiwan cancer Statistics show that lung cancer mortality rate are the highest among all other cancer types. Three main types of lung cancer have been discovered: squamous lung cancer, small cell lung cancer and non-small cell lung cancer (NSCLC). Above all three types of lung cancer, 85% belongs to non-small cell lung cancers. Lung adenocarcinoma is the most common type of NSCLC. Many factors are known to increase the risk of cancer, including environmental toxins, oncogene mutation or inheritance, etc.
The first part of our research is to investigate the effect of phthalates in lung adenocarcinoma cells. Phthalates (DEHP) incident in Taiwan was one of the most serious food safety issues that ever occurred. Exposure of the DEHP has been suggested to increase malignancy of many cancers in animal studies. But the relationship between DEHP and lung cancer was still unclear. According to our findings. Chronic DEHP exposure can enhance migration ability in CL1-0. Microarray analysis revealed that DEHP exposure influenced many genes related to cell metastasis、cell-adhesion and cell proliferation. We identified one important gene, anterior gradient homolog 2 (AGR2) that has been reported to play crucial roles in metastatic progression of cancer cells which might contribute to the mechanisms of DEHP-induced tumor progression. Herein, we provide possible mechanisms revealing how chronic DEHP exposure modulates lung adenocarcinoma progression. The second part of this study is to identify for potential metabolite biomarkers of lung cancer with EGFR mutations. L858R mutation in exon 21 of EGFR is the most common mutation in NSCLC. Lung cancer patients with activating EGFR mutations usually receive long term tyrosine kinase inhibitors treatment. However, a secondary mutation of T790M in EGFR would cause resistance to TKI treatment which leads to bad prognosis. Therefore, development of effective molecular biomarkers for early diagnosis becomes increasingly important in NSCLC patients. Transgenic engineered mice with Tet-on human EGFR T790M+L858R or EGFR L858R system were used to conditionally develop lung adenocarcinomas. Twenty metabolites were identified in lung tissue of EGFR mutant mice involving several key metabolic pathways such as glycolysis, tricarboxylic acid (TCA) cycle, fatty acids and methionine-salvage metabolism. Our metabolite profiling data also reveals several key metabolites such as spermine and hypoxanthine which are crucial in tumor progressions. Taken together, findings of this study suggest that DEHP might play crucial roles in lung adenocarcinoma progression. Furthermore, metabolite profile of lung cancer with EGFR mutation may provide potential therapeutic targets for lung cancer treatments. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T00:14:31Z (GMT). No. of bitstreams: 1 ntu-102-R00447006-1.pdf: 3312008 bytes, checksum: 5ec663a3f0426443d25158611ac2eeca (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 目錄(Contents)
口試委員審定書 謝誌 中文摘要 i 英文摘要 ii 目錄 iv 圖目錄 vii 表目錄 viii 縮寫表 ix Part 1 鄰苯二甲酸二(2-乙基己基)酯對於肺腺癌之癌化過程影響探討 Effects of Di-(2-ethylhexyl) Phthalate on Tumor Progression of Lung Adenocarcinoma Cells 中文摘要 2 英文摘要 3 第一章 緒論(Introduction) 5 1.1鄰苯二甲酸二(2-乙基己基)酯 (Di-(2-ethylhexyl) Phthalate, DEHP) 6 1.2鄰苯二甲酸二(2-乙基己基)酯 ( DEHP ) 毒理背景介紹 7 1.3鄰苯二甲酸二(2-乙基己基)酯 ( DEHP ) 與癌症之關聯 8 1.4 DEHP與肺相關疾病之關聯 9 1.5研究動機 9 第二章 材料與方法(Materials and Methods) 11 2.1實驗材料 12 2.1.1細胞株(Cell lines) 12 2.1.2藥品與試劑(Chemicals and Reagents) 12 2.2實驗方法 13 2.2.1細胞株(Cell lines)培養 13 2.2.2細胞毒性測試 (Cell viability test /MTT assay) 14 2.2.3細胞群落形成分析(Colony formation assay) 14 2.2.4細胞穿透試驗(Transwell assay) 14 2.2.5西方墨點法(Western blot analysis) 15 2.2.6核醣核酸之萃取 (RNA isolation)與cDNA之製備 15 2.2.7免疫螢光染色 16 2.2.8 cDNA微陣列(cDNA microarray) 16 2.2.9即時聚合酶鏈鎖反應(Real-time polymerase chain reaction) 16 2.2.10建立質體 16 2.2.11質體轉殖(Plasmid transfection) 17 2.2.12 AGR2基因拷貝數分析 18 2.3統計分析(Statistic analysis) 18 第三章 實驗結果 (Results) 19 3.1暴露DEHP對人類肺腺癌細胞株之存活率無顯著影響 20 3.2長期暴露DEHP對於人類支氣管上皮細胞之轉型無顯著影響 20 3.3長期暴露DEHP對於肺腺癌細胞株之抗藥性影響無顯著差異 20 3.4長期暴露DEHP會造成人類肺腺癌細胞型態改變以及增加轉移能力 21 3.5利用DNA微陣列分析長期暴露DEHP之人類肺腺癌細胞株 21 3.6長期暴露DEHP會促使人類肺腺癌細胞株CL1-0以及CL1-5之AGR2表現量上升 22 3.7移除DEHP後並不影響AGR2的表現量 23 3.8長期暴露DEHP造成AGR2表現量上升的可能機轉 23 3.9建立高表現AGR2的CL1-0細胞株 23 3.10高表現AGR2蛋白量的CL1-0其細胞轉移能力較高 23 第四章 討論(Discussion) 24 圖表集 (Figure and Tables) 29 Part 2以代謝體學探討上皮細胞生長因子接受器突變在肺癌中之角色 Metabonomic Study of Lung Adenocarcinoma with EGFR Mutation 中文摘要 60 英文摘要 61 第一章 緒論(Introduction) 62 1.1肺癌 63 1.2上皮細胞生長因子接受器(epidermal growth factor receptor, EGFR)與癌症 63 1.3 EGFR的突變 63 1.4代謝體學及於癌症研究之應用 64 1.5帶有突變EGFR基因轉殖小鼠模式 65 1.6研究動機 65 第二章 材料與方法(Materials and Methods) 67 2.1實驗材料 68 2.1.1動物模式 68 2.1.2藥品(Chemicals) 68 2.2實驗方法 68 2.2.1樣品的製備 68 2.2.2 Ultra High Pressure Liquid Chromatography (UHPLC) 69 2.2.3 Mass Spectrometry (MS) 69 2.2.4 UHPLC-MS Data analysis 69 2.3統計方法 69 第三章 實驗結果(Results) 70 3.1肺組織中具顯著差異之代謝物 71 3.2肺組織中影響的代謝途徑 71 第四章 討論(Discussion) 73 圖表集 (Figure and Tables) 78 總結(Conclusion) 88 參考文獻(Reference) 90 附錄 1 鄰苯二甲酸酯 102 附錄 2 pcDNA3.1(+)-AGR2質體之圖譜 103 | |
dc.language.iso | zh-TW | |
dc.title | 鄰苯二甲酸二(2-乙基己基)酯對於肺腺癌之癌化過程影響探討 | zh_TW |
dc.title | Effects of Di-(2-ethylhexyl) Phthalate on Tumor Progression of Lung Adenocarcinoma Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 郭錦樺,陳健尉,蘇剛毅 | |
dc.subject.keyword | 鄰苯二甲酸二(2-乙基己基)酯,肺腺癌,上皮生長因子,代謝體, | zh_TW |
dc.subject.keyword | Di-(2-ethylhexyl) Phthalate,Lung Adenocarcinoma,Epidermal growth factor receptor,Metabolomics, | en |
dc.relation.page | 103 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2013-07-31 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 毒理學研究所 | zh_TW |
顯示於系所單位: | 毒理學研究所 |
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