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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 羅凱尹(Kai-Yin Lo) | |
| dc.contributor.author | Yi-Min Huang | en |
| dc.contributor.author | 黃詒敏 | zh_TW |
| dc.date.accessioned | 2023-03-19T22:31:20Z | - |
| dc.date.copyright | 2022-08-29 | |
| dc.date.issued | 2022 | |
| dc.date.submitted | 2022-08-25 | |
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BioMed Research International. 2020:5205142 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/84894 | - |
| dc.description.abstract | 稀土元素鑭(Lanthanum)和鈰(Cerium)，其物理特性皆與鈣(Calcium)相似，若使用恰當，普遍被認為無毒性。而我們於前人的研究中發現鑭離子具有抗腫瘤調節的能力，因癌細胞內的酸性環境可助於的鑭及鈰解離，並以模擬鈣的形式干擾細胞生長。而本研究選用三陰性的乳腺癌細胞系MDA-MB-231，此種乳腺癌容易早期轉移，只能透過化療治癒。因此，我們透過檢測與核醣體生合成相關之因素及氧化壓力所造成的細胞損害以探討鑭及鈰是否會影響MDA-MB-231生長及其他造成毒性的機制。實驗結果顯示鑭及鈰會干擾rRNA的加工和核醣體生合成，其中也發現鑭及鈰對核醣體大次單元的影響更大，並且造成p53依賴性及非依賴性的核仁壓力。鑭及鈰的處理下也會誘導氧化壓力造成粒線體功能障礙及導致蛋白質氧化，而這些氧化的蛋白質可能因蛋白酶體無法發揮功能而在細胞中累積並誘導自噬作用。而由次世代定序的結果也發現細胞骨架相關基因受到影響，透過免疫螢光實驗可見到細胞經處理後導致F-actin聚集，表示細胞貼附可能受到影響。另外，我們也發現順鉑和鑭或鈰的處理下能產生加成性，抑制MDA-MB-231的生長及抗藥性相關的基因。本研究探討類鈣元素鑭及鈰抑制乳腺癌細胞生長之機制，其中核仁壓力及氧化壓力為造成細胞生長抑制之主因。 | zh_TW |
| dc.description.abstract | The rare earth elements lanthanum (La) and cerium (Ce) have similar physical properties to calcium (Ca) and are generally considered non-toxic if appropriately used. But La and Ce ions have the anti-tumor ability because the acidic environment in cancer cells can help the dissociation of La and Ce and interfere with cell growth in competition with Ca. In this study, the triple-negative breast cancer cell line MDA-MB-231 was selected, which is prone to early metastasis and can only be cured by chemotherapy. Therefore, we investigated a mechanism study about how La and Ce affect MDA-MB-231. The results show that La and Ce interfere with rRNA processing and ribosome biosynthesis and cause p53-dependent and independent nucleolar stress. La and Ce also induce oxidative stress through mitochondrial dysfunction, leading to protein oxidation. Autophagy is induced while the proteasome function is also impaired. The results of Next-Generation Sequencing (NGS) also found that cytoskeleton-related genes were affected. Immunofluorescence experiments showed that F-actin aggregated after cells were treated, indicating that cell adhesion may be affected. In addition, the co-treatment of cisplatin with La or Ce showed synergistic inhibition of growth and drug resistance-related genes in MDA-MB-231 cells. This study investigated the mechanism of the calcium-like elements La and Ce in inhibiting the growth of breast cancer cells. We found that nucleolar stress and oxidative stress were the main reasons for cell growth inhibition. | en |
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| dc.description.tableofcontents | 目錄 致謝 ii Abstract iii 摘要 iv 表目錄 ix 一、文獻回顧 1 1.1稀土元素之介紹 1 1.2鑭的基本性質與毒性探討 3 1.3鈰的基本性質與毒性探討 4 1.4鈰與鑭的藥理應用 6 1.5核醣體與核醣體生合成 7 1.6核仁壓力(Nucleolar stress) 8 二、研究動機 11 三、材料與方法 12 3.1細胞培養 12 3.2鑭離子金屬溶液配置 12 3.3鈰離子金屬溶液配置 12 3.4細胞活性測試 (MTT assay) 12 3.4.1 MTT reagent溶液配置： 13 3.4.2實驗步驟： 13 3.5細胞週期分析 13 3.6 Annexin V assay 14 3.7 西方墨點法(Western blotting) 14 3.7.1 收取蛋白質樣品： 14 3.7.2 蛋白質樣品濃度之測定及樣品製備： 14 3.7.3 SDS-PAGE膠體配置： 15 3.7.4 Western blotting： 15 3.8 免疫螢光染色(Immunofluorescence) 15 3.9 氧化壓力指標測定 16 3.10 蛋白質羰基化 (carbonylation)測定 16 3.10.1收取蛋白質樣品： 16 3.10.2蛋白質樣品濃度之測定及定量： 16 3.10.2羰基化蛋白質之測定及定量： 17 3.11觀察粒線體膜電位及型態 17 3.12 qPCR基因表達分析 17 3.12.1 cDNA樣品製備： 17 3.12.2即時定量聚合酶連鎖反應： 18 3.13 核醣體次單元圖譜分析 18 3.14多核醣體圖譜分析 19 3.15 BrdU incorporation assay 19 3.16北方墨點法 19 3.16.1 RNA萃取： 19 3.16.2 甲醛凝膠電泳分離rRNA 20 3.16.3 探針雜合及顯影： 20 3.17次世代基因定序 21 3.17.1 RNA萃取： 21 3.17.2轉錄體基因測序： 21 3.17.3生物資訊分析： 21 3.18統計分析 22 四、結果 23 4.1 金屬鑭、鈰使MDA-MB-231存活率下降 23 4.2金屬鑭、鈰使MDA-MB-231細胞週期停滯 24 4.3金屬鑭、鈰不誘導MDA-MB-231細胞凋亡 25 4.4金屬鑭及鈰使MDA-MB-231產生氧化壓力 28 4.5金屬鑭及鈰誘發MDA-MB-231產生核仁壓力 30 4.6 金屬鑭及鈰減少MDA-MB-231中核醣體大次單元的生合成 31 4.7金屬鑭及鈰使MDA-MB-231的rRNA裁切過程產生問題 32 4.8金屬鑭及鈰抑制MDA-MB-231的轉譯作用 33 4.9金屬鑭及鈰影響MDA-MB-231的基因表達 34 4.10 金屬鑭及鈰影響MDA-MB-231細胞骨架的完整性 36 4.11金屬鑭及鈰抑制MDA-MB-231細胞的蛋白酶體活性、誘導自噬作用 37 4.12 金屬鑭及鈰和順鉑共處理可使MDA-MB-231細胞存活率下降 39 4.13 金屬鑭及鈰和順鉑共處理可使MDA-MB-231細胞週期停滯 40 4.14 金屬鑭及鈰和順鉑共處理可誘導MDA-MB-231細胞凋亡 41 4.15 金屬鑭或鈰和順鉑共處理可抑制MDA-MB-231之PI3K/Akt路徑的活化 42 4.16 MDA-MB-231經金屬鑭或鈰和順鉑共處理後使NPM1易位 43 4.17金屬鑭或鈰和順鉑共處理影響細胞貼附 44 五、討論 45 5.1鑭及鈰可能藉由與鈣離子的相似性以影響核醣體及蛋白酶體 45 5.2鑭及鈰處理對核仁蛋白NPM1的影響 47 5.3鑭或鈰離子可能誘導失巢凋亡 (anoikis) 49 5.4 鑭或鈰可能藉由與鈣的相似性影響MDA-MB-231的細胞週期 51 六、結論 53 七、參考文獻 54 八.附錄 89 圖目錄 Figure 1. 金屬鑭、鈰使MDA-MB-231存活率下降 66 Figure 2. 金屬鑭、鈰使MDA-MB-231細胞週期停滯 67 Figure 3. 金屬鑭、鈰不誘導MDA-MB-231細胞凋亡 69 Figure 4. 金屬鑭及鈰使MDA-MB-231產生氧化壓力 71 Figure 5. 金屬鑭及鈰誘發MDA-MB-231產生核仁壓力 72 Figure 6. 金屬鑭及鈰減少MDA-MB-231中核醣體大次單元的生合成 73 Figure 7. 金屬鑭及鈰使MDA-MB-231的rRNA裁切過程產生問題 74 Figure 8. 金屬鑭及鈰抑制MDA-MB-231的轉譯作用 75 Figure 9. 金屬鑭及鈰影響MDA-MB-231的基因表達 80 Figure 10. 金屬鑭及鈰影響MDA-MB-231細胞骨架的完整性 81 Figure 11. 金屬鑭及鈰抑制MDA-MB-231細胞的蛋白酶體活性、誘導自噬作用 82 Figure 12. 金屬鑭及鈰和順鉑共處理可使MDA-MB-231細胞存活率下降 83 Figure 13. 金屬鑭及鈰和順鉑共處理可使MDA-MB-231細胞週期停滯 84 Figure 14. 金屬鑭及鈰和順鉑共處理可誘導MDA-MB-231細胞凋亡 85 Figure 15. 金屬鑭或鈰和順鉑共處理可抑制MDA-MB-231之PI3K/Akt路徑的活化 86 Figure 16. MDA-MB-231經金屬鑭或鈰和順鉑共處理後使NPM1易位 87 Figure 17. 金屬鑭或鈰和順鉑共處理影響MDA-MB-231貼附 88 附圖一、核醣體生合成示意圖(Pelletier et al., 2018) 89 附圖二、細胞凋亡之內在途徑及外在途徑(Taylor et al., 2008) 90 附圖三、泛素蛋白酶體系統及自噬作用示意圖(Pohl and Dikic, 2019) 91 附圖四、細胞週期中所需之鈣離子含量示意圖(Pande et al., 1996) 92 表目錄 Table 1. 本研究使用之一級抗體 62 Table 2. 本研究使用之二級抗體 62 Table 3. 研究中使用之北方墨點法探針序列 63 | |
| dc.language.iso | zh-TW | |
| dc.subject | 自噬作用 | zh_TW |
| dc.subject | 稀土元素 | zh_TW |
| dc.subject | 鑭 | zh_TW |
| dc.subject | 鈰 | zh_TW |
| dc.subject | 核醣體生合成 | zh_TW |
| dc.subject | 氧化壓力 | zh_TW |
| dc.subject | oxidative stress | en |
| dc.subject | Autophagy | en |
| dc.subject | Rare earth elements | en |
| dc.subject | Lanthanum | en |
| dc.subject | Cerium | en |
| dc.subject | Ribosome biogenesis | en |
| dc.title | 鑭及鈰抑制乳癌細胞生長之機制探討 | zh_TW |
| dc.title | Mechanism study of how lanthanum and cerium block the growth of breast cancer cell | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 110-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 謝佳倩(Chia-Chien Hsieh),楊珺堯(Chun-Yao Yang),楊雅雯(Ya-Wen Yang) | |
| dc.subject.keyword | 稀土元素,鑭,鈰,核醣體生合成,氧化壓力,自噬作用, | zh_TW |
| dc.subject.keyword | Rare earth elements,Lanthanum,Cerium,Ribosome biogenesis,oxidative stress,Autophagy, | en |
| dc.relation.page | 92 | |
| dc.identifier.doi | 10.6342/NTU202202791 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2022-08-26 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 農業化學研究所 | zh_TW |
| dc.date.embargo-lift | 2022-08-29 | - |
| 顯示於系所單位： | 農業化學系 | |
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