Solasodine，BP010W成份之一，抑制肺癌的遷移和侵襲能力

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屠少華(Shao-Hua Tu) 查詢紙本館藏

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系統生物與生物資訊研究所

論文名稱

Solasodine，BP010W成份之一，抑制肺癌的遷移和侵襲能力
(Solasodine, One of BP010W Component, Inhibits Migration and Invasion Ability in Lung Cancer)

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摘要(中)

肺癌具有很高的死亡率和和很低的治癒率，其中最重要的是轉移率非常的高，而放射治療和化學治療都會產生許多的副作用，目前常用於治療肺癌的標靶治療，後期也會產生強烈的抗藥性，因此，我們希望可以找到新的治療，而許多文獻顯示天然藥物可以用來治療癌症，除了毒性較低也會有較低的副作用。經過藥物的篩選找到了中藥BP010W可以抑制肺癌細胞的爬行能力，因此本文選擇BP010W做更深入的研究。我們將BP010W藥物分層，依據乙醇濃度變化量0%、20%、50%和95%，依序得到不同的成分，並發現BP010W 20%和BP010W 50%的分離物有更好的抑制效果。我們也透過利用生物資訊的方法和文獻的參考探討雌激素途徑會影響肺癌的調控，因此我們篩選出中藥BP010W裡的類固醇生物鹼Solasodine，利用HPLC分析顯示Solasodine出現在我們的BP010W 20%和BP010W 50%的分離物中，也經過加藥實驗證明Solasodine會抑制肺癌細胞的爬行能力，接著也利用西方墨點法，證明爬行相關的蛋白被抑制。最後利用爬行實驗證明了雌激素會增加肺癌細胞的爬行能力，我們也將Solasodine和雌激素共同治療，也發現彼此能互相抗衡爬行的能力。所以我們認為Solasodine可能可以當作新的肺癌治療候選藥物。

摘要(英)

Lung cancer has a high mortality rate and a very low cure rate. The most important one is that the Metastasis rate is very high, and both radiotherapy and chemotherapy have many side effects. Currently, it is often used for the Target treatment of lung cancer. It produces strong resistance, so we hope to find new treatments, and many literatures show that natural medicines can be used to treat cancer, with lower toxicity and lower side effects. After drug screening, it was found that Traditional Chinese Medicine BP010W can inhibit the migration ability of lung cancer cells., thus was selected for further research. The column extraction was performed for isolation of prue compounds from BP010W using different concentrations of ethanol, 0%, 20%, 50% and 95%, and we found the concentrations of 20% and 50% have most effective in inhibiting migration of cell. We assay the bioinformatics and literature references, and It is concluded that the estrogen pathway affects the regulation of lung cancer. Therefore, we screened out the steroid alkaloid Solasodine in Chinese medicine BP010W. HPLC analysis showed that Solasodine appeared in our BP010W 20% and BP010W 50% isolates. It was also confirmed by dosing experiments that Solasodine inhibited the migration ability of lung cancer cells, and then used Western blotting to prove that the migration-related proteins were inhibited. Finally, we using the wound-Healing assay to prove that estrogen will increase the migration ability of lung cancer cells, we also treated Solasodine and estrogen together, and found that each other can compete with each other.

關鍵字(中)

★ 肺癌
★ 轉移
★ 中草藥
★ 龍葵
★ 雌激素

關鍵字(英)

★ LL/2(LLC)
★ Solasodine

論文目次

中文摘要 I
Abstract II
圖目錄 V
表目錄 V
補充圖目錄 VIII
補充表目錄 IX
緒論 1
1-1 肺癌 1
1-2 肺癌轉移與治療 3
實驗方法與材料方法 6
2-1 細胞培養 6
2-2 藥物製備 6
2-3 傳統中藥的萃取 7
2-4 Wound-healing assay 細胞爬行實驗 8
2-5 Transwell-assay 細胞侵襲實驗 8
2-6 藥物分離 9
2-7 藥物接合模擬 10
2-8 晶片分析 10
2-9 高效液相色譜 11
2-10 蛋白質提取 11
2-11 西方墨點法 12
2-12 質粒提取 12
2-13 轉染 13
2-14 螢光素酶測定 13
結果 14
3-1 BP010W不同濃度對細胞爬行能力 14
3-1-1 Wound-healing 分析細胞爬行能力 14
3-1-2 Transwell-migration 分析細胞爬行能力 15
3-2 BP010W分離物對細胞爬行能力的影響 16
3-2-1 BP010W逆向管柱層析分離 16
3-2-2 Wound-healing 分析細胞爬行能力 17
3-2-3 Transwell分析細胞爬行能力 18
3-3 生物資訊分析 20
4-3-1 生物晶片 20
3-3-2 晶片分析 21
3-3-3 雌激素信號途徑相關基因的表現 22
3-4 BP010W中有效成分的篩選 27
3-4-1 藥物對接模擬 27
3-4-2 高效液相色譜分析BP010W中成份 32
3-5 Solasodine細胞學實驗 33
3-5-1 Wound-healing分析細胞爬行能力 33
3-5-2 Transwell分析細胞爬行能力 34
3-5-3 西方墨點法分析爬行相關的蛋白質表現 35
3-6 雌激素促進LLC細胞的爬行 37
3-6-1 Wound-healing分析細胞爬行能力 37
3-6-2 螢光素酶測定 38
討論 39
參考文獻 41
補充資料 46

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指導教授

蘇立仁(Li-Jen Su)

審核日期

2018-10-4

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