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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 
英文關鍵字 TARBP2  breast cancer  tamoxifen resistance 
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。
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.
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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