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| 系統識別號 |
U0026-0209201918205300 |
| 論文名稱(中文) |
以TARBP2為標的篩選在tamoxifen治療下對乳癌細胞具致敏性之中草藥萃取物 |
| 論文名稱(英文) |
The TARBP2-targeting drug screening identifies Chinese herbs that sensitize breast cancer cells to tamoxifen treatment |
| 校院名稱 |
成功大學 |
| 系所名稱(中) |
醫學檢驗生物技術學系 |
| 系所名稱(英) |
Department of Medical Laboratory Science and Biotechnology |
| 學年度 |
107 |
| 學期 |
2 |
| 出版年 |
108 |
| 研究生(中文) |
陳曦 |
| 研究生(英文) |
Hsi Chen |
| 學號 |
T36041065 |
| 學位類別 |
碩士 |
| 語文別 |
中文 |
| 論文頁數 |
40頁 |
| 口試委員 |
指導教授-陳百昇
口試委員-張雋曦
口試委員-邱慶豐
|
| 中文關鍵字 |
TARBP2
乳癌細胞
tamoxifen抗藥性
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| 英文關鍵字 |
TARBP2
breast cancer
tamoxifen resistance
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| 學科別分類 |
|
| 中文摘要 |
乳癌在全世界女性中是最常罹患的癌症。大約有70 ％的乳癌患者皆表達estrogen receptor (簡稱ER＋)。對於這一類的患者在臨床上常用的第一線用藥為tamoxifen。Tamoxifen主要藉由和雌激素(E2)競爭ER的接合，來阻止雌激素誘導癌細胞的生長。然而大約有5％的乳癌患者用tamoxifen治療5年後產生抗藥性。我們先前的研究發現用tamoxifen治療會誘導TAR RNA binding protein (TARBP2)的表達，從而使ER＋乳癌細胞降低對tamoxifen的敏感度。因此，本研究篩選來自陽明大學傳統醫藥研究所的81種中草藥萃取物，研究它們對tamoxifen誘導增加的TARBP2是否具有抑制效果，期能應用於合併治療。首先使用MTT assay確定單純加入中草藥萃取物，未對乳癌細胞(MCF-7)產生毒性。而在這些中草藥萃取物中，編號122-2和629的萃取物可顯著抑制因tamoxifen誘導的TARBP2蛋白質表現。此外還觀察到122-2和629提高乳癌細胞對tamoxifen的敏感度，並抑制因tamoxifen促進的TARBP2和SOX2蛋白質表現量。而在另一種ER＋乳癌細胞ZR75-1也觀察到類似的結果。先前研究結果顯示，p-AKT會正向調控TARBP2的蛋白質表現量，我們也證明這兩種中草藥藉由抑制p-AKT的表現量促使Merlin表現量增加，進而抑制TARBP2的蛋白質表現量。總結來說，我們鑑定出2種具有潛在性中草藥萃取物可增加乳癌細胞對tamoxifen的敏感度。
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| 英文摘要 |
Breast cancer is the most common cancer in women worldwide. Approximately 70% of breast cancer patients are estrogen receptor expression positive (ER+). One of the most commonly used first-line therapies for ER+ breast cancer patients is tamoxifen. Tamoxifen inhibits estrogen-induced cell growth predominately by competitive blockade of the estrogen receptor. Unfortunately, about 5% of the breast cancer patients showed drug-resistance after tamoxifen treatment for five years. Our previous study demonstrated that tamoxifen induces TAR RNA-binding protein (TARBP2) expression to enhance drug resistance in ER+ breast cancer cells. In this study, we screened eighty-one herbal extracts from Institute of Traditional Medicine，National Yang-Ming University to study their effects on reducing tamoxifen-induced TARBP2 expression. We first used MTT assay to ensure that no cytotoxic effects was observed in herbal extract treatment. Among these extracts, 122-2 and 629, significantly inhibited tamoxifen-induced TARBP2 protein expression. Furthermore, we observed that either 122-2 or 629 enhances tamoxifen sensitivity and prevents tamoxifen-induced TARBP2 and SOX2 protein expression. Similar results were also observed in another ER+ breast cancer cell line, ZR75-1Previous study demonstrated that p-AKT downregulate TARBP2 protein level. We also found that either 122-2 or 629 prevents p-AKT protein expression and induces Merlin protein expression. Taken together, we identified two herbal extracts with potential in combination therapy which sensitize breast cancer cells to tamoxifen treatment.
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| 論文目次 |
目錄
中文摘要 I
Abstract II
致謝 Ⅵ
目錄 Ⅶ
Figure 索引 Ⅹ
縮寫表 Ⅺ
Chapter 1. 引言 1
1.1 乳癌 1
1.2 Tamoxifen 1
1.3 中草藥在癌症治療的應用 2
1.4 TARBP2 3
1.5聯合治療 4
Chapter 2. 材料與方法 6
2.1 材料 6
2.2.1細胞培養 7
2.2.2西方墨點法 7
2.2.3RNA萃取和即時定量聚合酶反應 8
2.2.4細胞感受性測驗 8
2.2.5利用plasmid過度表現TARBP2 9
2.2.6 Combination Index (CI)計算 9
2.2.7統計分析 10
Chapter 3. 實驗結果 11
3.1 單一中草藥萃取物對乳癌細胞之毒性測試 11
3.2 中草藥萃取物對因tamoxifen治療下誘發TARBP2蛋白質表現量的影響 11
3.3 TARBP2蛋白質表現量下降是否隨中草藥濃度增加而具有劑量依賴性 12
3.4 中草藥萃取物並非透過RNA層面調控TARBP2表現 12
3.5 MCF-7乳癌細胞於單一或聯合藥物治療下細胞存活率的比較 13
3.6 中草藥萃取物增加乳癌細胞對tamoxifen的敏感度是藉由抑制TARBP2表現量所導致 13
3.7 Tamoxifen所促進的TARBP2蛋白質穩定性是否受到中草藥萃取物治療的抑制 14
3.8 驗證不同來源之中草藥萃取物是否影響乳癌細胞對tamoxifen的敏感度 14
3.9 中草藥是否也增加另一株ER+乳癌細胞對tamoxifen的敏感性 15
3.10中草藥萃取物對tamoxifen所誘導p-AKT、抑制Merlin蛋白質表現量的影響 16
Chapter 4. 討論 17
References 19
Figures 24
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