- 學位論文
苧麻根萃取物反轉上皮間質轉化和抑制腫瘤細胞中非固著生長
Boehmeria Nivea Extract (BNE-RRC) Reverses Epithelial-Mesenchymal Transition and Inhibits Anchorage-Independent Growth in Tumor Cells
摘要
近年來表皮-間質細胞轉化（epithelial to mesenchymal transition, EMT）表型的出現被認為可能代表著使癌細胞獲得與惡性腫瘤相關的特性。EMT其實在生物學上也可以發現到的動態過程，在正常織器官的發育就會發現他的參與，其中上皮細胞失去其上皮特徵並獲得間質的轉變。而在生理上，EMT 在重塑、促進組織癒合、修復以及應對各種組織損傷的纖維化方面也發揮著重要作用。在癌症中，EMT也與腫瘤所使用的存活策略經由侵襲、起始、轉移等來增加存活，以便於來拓展生存範圍和對化療藥物的抵抗增加都可能有密切的關聯性。 苧麻根萃取物，經證實無毒性且於B型肝炎造成的肝病療效良好，也有苧麻根萃取物相關試驗抗發炎的功效，本研究也將針對苧麻根萃取物對於口腔鱗狀細胞株-KB細胞株進行有關EMT標誌蛋白的表達進一步探討。 實驗方法主要是以下列的方式來測試KB細胞經苧麻根萃取物處理之後在對照組和實驗組之間各種表現差異：1.軟瓊脂非固著生長測試，2.以西方墨點法確認EMT 標誌蛋白質表達量，3. KB細胞在經苧麻根萃取物作用下對於migration 、invasion和wound healing assay 會造成抑制 ，4.對化療藥物降低其阻抗的表現。結果:在不固著生長方面，苧麻根萃取物可以有效抑制癌細胞系中非固著生長。及抑制 EMT 標誌物vimentin，desmin的表達明顯下調，並增加表皮-間質細胞轉型(MET)標誌物α-catenin的表達量。苧麻根萃取物可以有效抑制癌細胞的生長、migration和invasion以及上皮形態的誘導，更有助於增加對化療藥doxorubicin的敏感性。 KB細胞在苧麻根萃取物的長時間處理實驗中，發現可以在抑制 EMT 和隨後觸發 MET 方面發揮不可或缺的作用。 此外，用苧麻根萃取物處理腫瘤細胞 14天顯示間質標誌物vimentin的表達降低； 即使此後苧麻根萃取物在培養中停用 4 週，這些細胞也表現出對化療藥物doxorubicin的敏感性增加。 因此，苧麻根萃取物具有促進腫瘤細胞分化和間質表型丟失(loss of mesenchymal phenotype)的巨大潛力。
並列摘要
Epithelial-Mesenchymal-transition (EMT) phenotype, which has emerged in recent years, may represent a potential clinical marker. EMT is a biological process in which cells lose their epithelial characteristics and acquire mesenchymal features. Physiologically, EMT plays an essential role in remodeling, promoting tissue healing, repairing as well as fibrosis in response to a variety of tissue damages. In cancer, EMT is associated with tumor invasion, initiation, metastasis and resistance to chemotherapy. Boehmeria Nivea Extract (BNE-RRC) is extracted from the root of Boehmeria nivea. It has been proved non-toxic and has a good effect for liver disease caused by hepatitis B. There have been BNE-RRC related tests about the anti-inflammatory efficacy. This study investigated BNE-RRC treatment oral squamous carcinoma KB cell line and tested, the expression of EMT marker protein. The experimental methods are as follows. 1. anchorage-independent growth test on soft agar 2. confirmation of EMT marker protein expression by Western blotting method 3. motility of KB cells in migration, invasion and wound healing assays. 4. performance of resistance to chemotherapy drugs. We report that Boehmeria Nivea Extract (BNE-RRC) could effectively inhibit anchorage-independent growth in cancer cell lines KB. Furthermore, root of Boehmeria nivea could suppress the expression of EMT markers, such as Vimentin and Desmin, and increase the expression of mesenchymal-epithelial-transition (MET) markerα-catenin. It is suggested that BNE-RRC could play an integral role towards inhibiting EMT and subsequently triggers MET. Moreover, treatments of tumor cells with BNE-RRC for a period of 14 days would decrease in the expression of mesenchymal marker, vimentin; even upon the withdrawal of BNE-RRC for 4 weeks thereafter in culture.Tthese cells demonstrated an increased sensitivity to chemotherapeutic drug, doxorubicin. Therefore, BNE-RRC possesses a great potential attributed to promoting tumor cell promoted differentiation and loss of mesenchymal phenotype. We report that Boehmeria Nivea Extract (BNE-RRC) could effectively inhibit anchorage-independent growth in cancer cell lines KB. Furthermore, BNE-101 could suppress the expression of EMT markers, such as vimentin and desmin, and increase the expression of mesenchymal-epithelial-transition (MET) marker α-catenin.