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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳彥榮 | zh_TW |
| dc.contributor.advisor | Edward Chern | en |
| dc.contributor.author | 邱鈺惠 | zh_TW |
| dc.contributor.author | Yu-Hui Chiu | en |
| dc.date.accessioned | 2021-07-10T21:51:10Z | - |
| dc.date.available | 2024-08-20 | - |
| dc.date.copyright | 2019-08-26 | - |
| dc.date.issued | 2019 | - |
| dc.date.submitted | 2002-01-01 | - |
| dc.identifier.citation | 1 Balkwill, F. R., Capasso, M. & Hagemann, T. The tumor microenvironment at a glance. J. Cell Sci. 125, 5591-5596, doi:10.1242/jcs.116392 (2012).
2 Chen, L., Endler, A. & Shibasaki, F. Hypoxia and angiogenesis: regulation of hypoxia-inducible factors via novel binding factors. Exp. Mol. Med. 41, 849-857, doi:10.3858/emm.2009.41.12.103 (2009). 3 Corbet, C. & Feron, O. Tumour acidosis: from the passenger to the driver's seat. Nature Reviews Cancer 17, 577, doi:10.1038/nrc.2017.77 (2017). 4 San-Millán, I. & Brooks, G. A. Reexamining cancer metabolism: lactate production for carcinogenesis could be the purpose and explanation of the Warburg Effect. Carcinogenesis 38, 119-133, doi:10.1093/carcin/bgw127 (2017). 5 Pugh, C. W. & Ratcliffe, P. J. Regulation of angiogenesis by hypoxia: role of the HIF system. Nat. Med. 9, 677-684, doi:10.1038/nm0603-677 (2003). 6 Krock, B. L., Skuli, N. & Simon, M. C. Hypoxia-induced angiogenesis: good and evil. Genes Cancer 2, 1117-1133, doi:10.1177/1947601911423654 (2011). 7 Karsch-Bluman, A. et al. Tissue necrosis and its role in cancer progression. Oncogene 38, 1920-1935, doi:10.1038/s41388-018-0555-y (2019). 8 Pollheimer, M. J. et al. Tumor necrosis is a new promising prognostic factor in colorectal cancer. Hum. Pathol. 41, 1749-1757, doi:https://doi.org/10.1016/j.humpath.2010.04.018 (2010). 9 Dvorak, H. F. Tumors: wounds that do not heal. Similarities between tumor stroma generation and wound healing. N. Engl. J. Med. 315, 1650-1659, doi:10.1056/nejm198612253152606 (1986). 10 Srikrishna, G. & Freeze, H. H. Endogenous Damage-Associated Molecular Pattern Molecules at the Crossroads of Inflammation and Cancer. Neoplasia 11, 615-628, doi:10.1593/neo.09284 (2009). 11 Yu, H., Pardoll, D. & Jove, R. STATs in cancer inflammation and immunity: a leading role for STAT3. Nat. Rev. Cancer 9, 798-809, doi:10.1038/nrc2734 (2009). 12 Ben-Neriah, Y. & Karin, M. Inflammation meets cancer, with NF-κB as the matchmaker. Nat. Immunol. 12, 715, doi:10.1038/ni.2060 (2011). 1 Balkwill, F. R., Capasso, M. & Hagemann, T. The tumor microenvironment at a glance. J. Cell Sci. 125, 5591-5596, doi:10.1242/jcs.116392 (2012). 2 Chen, L., Endler, A. & Shibasaki, F. Hypoxia and angiogenesis: regulation of hypoxia-inducible factors via novel binding factors. Exp. Mol. Med. 41, 849-857, doi:10.3858/emm.2009.41.12.103 (2009). 3 Corbet, C. & Feron, O. Tumour acidosis: from the passenger to the driver's seat. Nature Reviews Cancer 17, 577, doi:10.1038/nrc.2017.77 (2017). 4 San-Millán, I. & Brooks, G. A. Reexamining cancer metabolism: lactate production for carcinogenesis could be the purpose and explanation of the Warburg Effect. Carcinogenesis 38, 119-133, doi:10.1093/carcin/bgw127 (2017). 5 Pugh, C. W. & Ratcliffe, P. J. Regulation of angiogenesis by hypoxia: role of the HIF system. Nat. Med. 9, 677-684, doi:10.1038/nm0603-677 (2003). 6 Krock, B. L., Skuli, N. & Simon, M. C. Hypoxia-induced angiogenesis: good and evil. Genes Cancer 2, 1117-1133, doi:10.1177/1947601911423654 (2011). 7 Karsch-Bluman, A. et al. Tissue necrosis and its role in cancer progression. Oncogene 38, 1920-1935, doi:10.1038/s41388-018-0555-y (2019). 8 Pollheimer, M. J. et al. Tumor necrosis is a new promising prognostic factor in colorectal cancer. Hum. Pathol. 41, 1749-1757, doi:https://doi.org/10.1016/j.humpath.2010.04.018 (2010). 9 Dvorak, H. F. Tumors: wounds that do not heal. Similarities between tumor stroma generation and wound healing. N. Engl. J. Med. 315, 1650-1659, doi:10.1056/nejm198612253152606 (1986). 10 Srikrishna, G. & Freeze, H. H. Endogenous Damage-Associated Molecular Pattern Molecules at the Crossroads of Inflammation and Cancer. Neoplasia 11, 615-628, doi:10.1593/neo.09284 (2009). 11 Yu, H., Pardoll, D. & Jove, R. STATs in cancer inflammation and immunity: a leading role for STAT3. Nat. Rev. Cancer 9, 798-809, doi:10.1038/nrc2734 (2009). 12 Ben-Neriah, Y. & Karin, M. Inflammation meets cancer, with NF-κB as the matchmaker. Nat. 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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77213 | - |
| dc.description.abstract | 癌細胞的進程與其微環境密切相關，腫瘤組織快速生長的特性會導致腫瘤細胞容易遭遇到低氧、營養缺乏等逆境，造成細胞死亡。由於臨床上常可見到腫瘤組織的壞死現象，腫瘤微環境的壓力是有利於癌細胞存活，抑或是癌細胞先天上的生存弱點，值得深入研究。細胞壞死會釋出內部分子引起發炎反應，其中，損傷相關分子模式 (Damage-associated molecular patterns, DAMPs) 是一群內生型可引起免疫反應的誘導物。高遷移率族蛋白1 (high-mobility group box 1, HMGB1) 被認為是細胞死亡時主要釋放出來的損傷相關分子，並經常在癌症組織中存在。細胞外的 HMGB1 具有細胞激素與趨化因子等活性，故參與許多發炎相關疾病的調控；然而在癌症相關文獻中， HMGB1 卻扮演促癌與抑癌雙重角色，顯示其功能尚未完全明瞭。
在我的研究中，希望找出癌細胞周邊環境的 HMGB1 對其自身的影響。為了探討胞外 HMGB1 對癌細胞的效應，本研究利用大腸桿菌 RosettaTM (DE3) 建構 His-HMGB1 表達與純化系統，可獲得純度高之重組蛋白，同時也利用真核細胞 HEK293FT 過量表現方式取得含有HMGB1 蛋白質的細胞萃取物。實驗結果發現，胞外的 HMGB1 具有促進大腸癌細胞 NF-κB 及 Wnt 訊息的活性，然而卻會抑制癌細胞的爬行能力與球體形成能力。 HMGB1 對癌細胞的調控可能和其轉譯後修飾有關，也是造成現有文獻報導促癌、抑癌相互矛盾的原因之一，若能更加明白 HMGB1 蛋白的修飾情形，則有助於預測其功能，因而在癌症醫學上可以視其作用擬定最適合的治療方案。 | zh_TW |
| dc.description.abstract | The progression of cancer is profoundly affected by its microenvironment. As tumors are rapid growing mass, cells inside tumor bulk may encounter adversities such as hypoxia and nutrient deprivation, which may cause cell death. Whether stress-induced cell death is advantageous for cancer cell survival or harmful as the innate vulnerability is an issue worth studying. Clinically, necrosis is typically seen in cancer tissues. During necrosis, intracellular molecules are passively released outside,triggering inflammation. Damage-associated molecular patterns (DAMPs) are endogenous inducers of inflammation which vary in categories and functions. One of the DAMPs, high-mobility group box 1 (HMGB1) protein is thought to be the major factor released by necrotic cells. Extracellular HMGB1 possesses cytokine and chemokine activities, thus participating in inflammation-associated diseases. The research about HMGB1 in cancer, however, revealed both its pro-tumor and anti-tumor roles. Therefore, the function of HMGB1 still remains to be clarified.
In this study, I aim to investigate how HMGB1 in the extracellular environment affects cancer cell properties. His-HMGB1 was produced in RosettaTM (DE3) E. coli strain and purified through affinity chromatography. HMGB1-overexpressing HEK293FT cell line was also constructed to obtain HMGB1 of eukaryotic origin. Extracellular HMGB1 was shown to activate NF-κB and Wnt activity in colorectal cancer cells; nevertheless, HMGB1 treatment may inhibit cancer cell migration and sphereforming ability. The effect of HMGB1 on cancer cells seems to has a lot to do with its post-translational modifications, which possibly leads to the pro-tumor and anti-tumor inconsistency in experimental results. The understanding of HMGB1 protein modifications would help predict its function. As a consequence, depending on the function of HMGB1 in cancer tissues, a suitable treatment strategy could be determined to combat cancer. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-10T21:51:10Z (GMT). No. of bitstreams: 1 ntu-108-R06b22010-1.pdf: 4597904 bytes, checksum: b514781bb2bc56ae7f55cbb808fc8204 (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | 中文摘要 I
Abstract II 目錄 IV 英文縮寫對照表 VII 第一章 背景介紹 1 1.1 腫瘤微環境 (Tumor microenvironment) 1 1.2 發炎與癌症 2 1.3 損傷相關分子模式 (Damage-associated molecular pattern, DAMP) 3 1.4 高遷移率族蛋白1 (High-mobility group box 1, HMGB1) 4 1.5 結直腸癌 (Colorectal cancer) 7 1.6 癌症幹細胞 (Cancer stem cells, CSCs) 7 1.7 高遷移率族蛋白1與癌幹細胞 9 第二章 研究動機與目的 10 第三章 材料與方法 11 3.1 質體建構 (Plasmid construction) 11 3.2 組胺酸標籤之高遷移率族蛋白1 (His-HMGB1) 表現與純化 11 3.3 細胞培養 (Cell culture) 13 3.4 RNA抽取 13 3.5 反轉錄聚合酶鏈反應 (Reverse transcription PCR, RT-PCR) 13 3.6 定量即時聚合酶鏈反應 (Quantitative real-time PCR, qPCR) 14 3.7 蛋白質抽取與西方墨點法 (Western blot) 14 3.8 細胞轉染 (Cell transfection) 及凍融法獲取細胞萃取物 15 3.9 冷光素酶報導基因測定 (Luciferase reporter assay) 15 3.10 細胞爬行試驗 (Cell migration assay) 15 3.11 細胞球體形成試驗 (Sphere forming assay) 16 3.12 統計分析 16 第四章 研究結果 17 4.1 HMGB1 及其受體 (receptor) 於結直腸癌細胞株中的表現 17 4.2 決定能最佳生產 His-HMGB1 的菌株和誘導條件 17 4.3 His-HMGB1 於大腸桿菌系統的純化 18 4.4 His-HMGB1 具有引起細胞發炎的能力 18 4.5 His-HMGB1 可能促進 Wnt 訊息活性 18 4.6 His-HMGB1 可能促進癌幹細胞相關基因的表現 19 4.7 His-HMGB1 降低結直腸癌細胞爬行能力與球體形成能力 19 4.8 以 HEK293FT 取得真核來源之 HMGB1 19 4.9 真核來源之 HMGB1 引起細胞發炎能力不顯著 20 4.10 真核來源之 HMGB1 可能促進 Wnt 訊息活性 20 4.11 真核來源之 HMGB1 降低細胞爬行與球體形成能力 21 第五章 討論與總結 22 5.1 大腸桿菌純化系統評估 22 5.2 HMGB1 自身引起發炎的能力可能不高 23 5.3 HMGB1 對 Wnt 訊息的影響 24 5.4 HMGB1 抑制大腸癌細胞惡化表徵 25 5.5 HMGB1 轉譯後修飾可能影響對癌細胞的功能 25 5.6 總結與展望 26 第六章 附表與結果圖 27 表一 使用引子列表 27 表二 使用抗體列表 28 圖一 HMGB1 及其受體於結直腸癌細胞株中的表現 29 圖二 決定最佳表現菌株及誘導條件優化 30 圖三 His-HMGB1 蛋白純化結果 33 圖四 His-HMGB1 提升 NF-κB 活性及發炎相關基因表現 34 圖五 His-HMGB1 提升 Wnt 活性及其下游基因表現 36 圖六 His-HMGB1 提升癌幹細胞相關基因表現 37 圖七 His-HMGB1 抑制細胞爬行能力與球體形成能力 39 圖八 真核來源之 HMGB1 可能提升 NF-κB 及 Wnt 活性 41 圖九 真核來源之 HMGB1 抑制細胞爬行能力與球體形成能力 42 第七章 參考文獻 43 附錄 期刊格式英文稿 i | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 損傷相關分子模式 | zh_TW |
| dc.subject | 高遷移率族蛋白1 | zh_TW |
| dc.subject | 發炎 | zh_TW |
| dc.subject | 腫瘤微環境 | zh_TW |
| dc.subject | Wnt 訊息 | zh_TW |
| dc.subject | 大腸癌 | zh_TW |
| dc.subject | colorectal cancer | en |
| dc.subject | tumor microenvironment | en |
| dc.subject | inflammation | en |
| dc.subject | damageassociated molecular pattern (DAMP) | en |
| dc.subject | high-mobility group box 1 (HMGB1) | en |
| dc.subject | Wnt signaling | en |
| dc.title | 胞外 HMGB1 於大腸癌細胞之角色探討 | zh_TW |
| dc.title | An investigation of the role of extracellular HMGB1 in colorectal cancer cells | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 107-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 廖憶純;黃楓婷 | zh_TW |
| dc.contributor.oralexamcommittee | Yi-Chun Liao;Feng-Ting Huang | en |
| dc.subject.keyword | 大腸癌,腫瘤微環境,發炎,損傷相關分子模式,高遷移率族蛋白1,Wnt 訊息, | zh_TW |
| dc.subject.keyword | colorectal cancer,tumor microenvironment,inflammation,damageassociated molecular pattern (DAMP),high-mobility group box 1 (HMGB1),Wnt signaling, | en |
| dc.relation.page | 69 | - |
| dc.identifier.doi | 10.6342/NTU201903668 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2019-08-16 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 生化科技學系 | - |
| 顯示於系所單位： | 生化科技學系 | |
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| ntu-107-2.pdf 未授權公開取用 | 4.49 MB | Adobe PDF |
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