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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/6503完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳兆勛 | |
| dc.contributor.author | Jie-Fu Chen | en |
| dc.contributor.author | 陳界甫 | zh_TW |
| dc.date.accessioned | 2021-05-17T09:14:14Z | - |
| dc.date.available | 2014-08-20 | |
| dc.date.available | 2021-05-17T09:14:14Z | - |
| dc.date.copyright | 2012-08-20 | |
| dc.date.issued | 2012 | |
| dc.date.submitted | 2012-08-16 | |
| dc.identifier.citation | [1] Vital Statistic: Taipei T, ROC: Department of Health, Executive Yuan,Health statistics of the Republic of China, 2005.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/6503 | - |
| dc.description.abstract | 肝癌在全世界是一種常見的癌症之一，在台灣也是導致許多癌症病患的死亡原因。為了提高肝癌病人的存活率，或降低手術過後的復發率，尋找有效的抗癌藥物是重要並且需要的。而廣泛存在於植物界的槲皮素因為具有抗癌變及抗增生的效果，且有強大的抗氧化能力以及在細胞訊號傳導、細胞週期、細胞凋亡的調控上有一定的能力而受到注意。本研究目的是希望藉由相關實驗，來觀察槲皮素是否能抑制人類肝癌細胞株Hep3B、Huh7，並以基因晶片做檢測分析，針對被調控到且會影響細胞增殖生長、細胞週期、細胞移動及細胞凋亡的基因做進一步探討，闡明槲皮素抑制人類肝癌細胞的藥理機制。此實驗以人類肝癌細胞株Hep3B及Huh7為材料進行體外培養，以各濃度槲皮素對細胞進行Proliferation、Colony formation、Wound-healing及Flow cytometry實驗，並以槲皮素影響效果較顯著的Huh7作進一步微陣列分析。實驗結果顯示，Hep3B及Huh7在48小時槲皮素濃度分別為500、50μM時，開始有顯著抑制細胞增殖效果；在濃度50、10μM時，顯著抑制細胞菌落發展；在同樣濃度100μM時，經過12小時就開始有顯著抑制細胞移動效果以及濃度分別在50、10μM能分別誘導Hep3B、Huh7產生細胞凋亡，且對細胞週期G1 phase都有阻滯效果，以上實驗效果都與槲皮素濃度成正相關；在來從進一步的Huh7微陣列分析實驗結果發現，槲皮素能調控Cell apoptosis、Cell cycle、Cell migration相關基因，而這些受到調控的基因可能參與了Apoptosis pathway、JAK/STAT、PI3K/AKT、MAPK/ERK pathway。而這些被影響調控的基因以及所參與的訊號傳導路徑就是槲皮素能夠抑制癌細胞生長增殖生長、細胞週期、細胞移動以及誘導凋亡背後最強而有力的證據。未來槲皮素或許可用於臨床上治療肝癌之輔助治療藥物，提高肝癌病人的存活率、降低化療藥物所產生的副作用以及降低手術過後的復發率。 | zh_TW |
| dc.description.abstract | Hetapocellular carcinoma is one of the common cancers in the world, it is also the main reason that makes people die in the cancer. To increase the survival rate or reduce the recurrence rate, it is necessary and important to find those efficacious anti-cancer drugs. Quercetin is common existing in plant with great effect of anti-carcinogenesis and anti- proliferation. It also has good capability on anti-oxidant. Besides, Quercetin has been noticed by the abilities on effecting cell signal transduction and regulating of cell cycle and inducing cell apoptosis. The purpose of this research is trying to observe whether Quercetin can inhibit HCC cell growth by several experiments. Moreover, to find that the mechanism of inhibiting the proliferation、growth、cell cycle、migration and inducing apoptosis on HCC cell by Quercetin, we used microarray so that we can analyze the suppression effect on HCC cell by gene expression. This experiment uses HCC cell line- Hep3 and Huh7 as the materials in vitro culture, and different concentrations of Quercetin are using on experiments of Proliferation, Colony formation, Wound-healing and Flow cytometry. Huh7 is used on microarray analysis due to the better influence on Quercetin as well. Experiment results show that Hep3B and Huh7 have significantly inhibiting proliferation when the concentration of Quercetin is 500, 50μM on 48 hours; it also shows inhibition on colony formation while the concentration is 50、10μM. And Quercetin has good inhibition effect on cell migration after 12 hours when the concentration is 100μM, and it can also induce Hep3B、Huh7 apoptosis amd arrest G1 phase in cell cycle when the concentration is 50、10μM. All effect by Quercetin during the experiments is depending on the concentrations of Quercetin. Moreover, the experiment of microarray analysis for Huh7 also shows that Quercetin can regulate the gene relative to Apoptosis、cell cycle and migration。And these gene regulated by Quercetin may be involve Apoptosis pathway and several signal transduction pathway：JAK/STAT、PI3K/AKT、MAPK/ERK pathway. All the effect on signal transduction pathway or gene regulated by Quercetin are the strong evidence that Quercetin can inhibit the growing of cancer cells、arrest cell cycle、suppress cell migration and induce the apoptosis of cancer cells. Perhaps people can use Quercetin on clinical medicine for curing liver cancer, increase the survival rate and also decrease the side effect and recurrence rate. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-17T09:14:14Z (GMT). No. of bitstreams: 1 ntu-101-R99543060-1.pdf: 4677545 bytes, checksum: 28b55d9ece120ccb3f7103222f00c095 (MD5) Previous issue date: 2012 | en |
| dc.description.tableofcontents | 第一章 前言 1
第二章 文獻回顧 3 一、 肝癌 3 二、 槲皮素 6 三、 細胞凋亡 10 四、 細胞週期 14 五、 基因晶片 17 第三章 材料和方法 20 一、 實驗材料 20 1. 細胞培養 20 2. 細胞計數 22 3. 藥物處理 23 4. Alamar Blue cell viability reagent 23 5. 流式細胞儀 25 6. AffymetrixR Human Genome U219 Array 26 二、 實驗方法 27 1. Cell proliferation assay 27 2. Colony formation assay 27 3. Wound-healing assay 28 4. Flow cytometry assay 28 5. Microarray analysis 29 第四章 結果 31 一、 Cell proliferation assay 31 二、 Colony formation assay 32 三、 Wound-healing assay 32 四、 Flow cytometry assay 33 五、 Microarray analysis 34 第五章 討論 36 第六章 結論 41 參考文獻 62 圖目錄 FIG. 1 Two major apoptotic pathways in mammalian cells. 12 FIG. 2人類肝癌細胞株 20 FIG. 3 血球計數器 22 FIG. 4 血球計數器上的方格 22 FIG. 5 Resazurin還原成Resorufin 24 FIG. 6 Resorufin的吸光度與放射螢光光譜 24 FIG. 7 AlamarBlue protocol 24 FIG. 8 細胞經雷射光激發產生的散射光及螢光 25 Fig. 9 槲皮素對Hep 3B的Proliferation assay結果 42 Fig. 10 槲皮素對Huh 7的Proliferation assay結果 43 Fig. 11 槲皮素對Hep 3B的Colony formation assay結果 44 Fig. 12 槲皮素對Huh 7的Colony formation assay結果 45 Fig. 13 槲皮素對Hep 3B的Wound-healing assay結果 46 Fig. 14 槲皮素對Huh 7的Wound-healing assay結果 47 Fig. 15 槲皮素對Hep 3B的Flow cytometry assay結果 48 Fig. 16 槲皮素對Huh 7的Flow cytometry assay結果 49 Fig. 17 RNA Electropherogram 50 Fig. 18 RNA Purity test 51 Fig. 19 aRNA Quality Control 51 Fig. 20 Microarray Image of Huh7-0μM 52 Fig. 21 Microarray Image of Huh7-50μM 53 FIG. 22槲皮素對肝癌調控兩倍以上基因之影響 61 表目錄 Table. 1 槲皮素上調2倍以上Apoptosis相關基因 54 Table. 2 槲皮素下調2倍以上Apoptosis相關基因 56 Table. 3 槲皮素上調2倍以上Cell cycle相關基因 57 Table. 4 槲皮素下調2倍以上Cell cycle相關基因 58 Table. 5 槲皮素上調2倍以上Migration相關基因 59 Table. 6 槲皮素下調2倍以上Migration相關基因 60 | |
| dc.language.iso | zh-TW | |
| dc.subject | 基因晶片 | zh_TW |
| dc.subject | 肝癌 | zh_TW |
| dc.subject | 槲皮素 | zh_TW |
| dc.subject | 細胞凋亡 | zh_TW |
| dc.subject | 細胞週期 | zh_TW |
| dc.subject | Quercetin | en |
| dc.subject | Hetapocellular carcinoma | en |
| dc.subject | Microarray | en |
| dc.subject | Cell cycle | en |
| dc.subject | Apoptosis | en |
| dc.title | 槲皮素對肝癌的影響 | zh_TW |
| dc.title | The effect of Quercetin in hepatocellular carcinoma | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 100-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 戴明泓,賴信志 | |
| dc.subject.keyword | 肝癌,槲皮素,細胞凋亡,細胞週期,基因晶片, | zh_TW |
| dc.subject.keyword | Hetapocellular carcinoma,Quercetin,Apoptosis,Cell cycle,Microarray, | en |
| dc.relation.page | 72 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2012-08-16 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 應用力學研究所 | zh_TW |
| 顯示於系所單位： | 應用力學研究所 | |
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|---|---|---|---|
| ntu-101-1.pdf | 4.57 MB | Adobe PDF | 檢視/開啟 |
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