- 學位論文
利用斑馬魚作為模式生物進行乳癌分析
Using zebrafish as an animal model for breast cancer analysis
摘要
近年來，因為斑馬魚飼養容易且成本較低，因此被廣泛應用於醫 學研究的模式生物，大多是用在胚胎發育、聽覺缺失及毒性分析。斑 馬魚在癌症研究上的應用目前大多集中在轉移及壓力承受分析，本研 究也大多會圍繞這些建立癌症的動物模式。另外，諸多研究顯示Hsp27 對於細胞的壓力承受能力有很大的關聯，對於調控細胞的抗凋亡、細 胞週期調控及治療抗性的相關機轉扮演了關鍵的角色，而下游機轉的 調控根據Hsp27 的磷酸化狀態改變所引起的Hsp27 蛋白聚合體構型的 不同而有所相異。綜合以上所述，本研究的目的主要利用斑馬魚模式 來探討Hsp27 對於乳癌轉移的影響。 本研究使用3 天大斑馬魚胚胎進行異種移植手術。而為了將 Hsp27 的磷酸化的因素納入實驗中，特別設計了Hsp27D(模擬磷酸化 活化型) 及Hsp27A( 模擬磷酸化死亡型) 兩種突變型並和 wtHsp27(Hsp27 過表現型)及控制組進行比較。在BT-474 乳癌細胞的 實驗中，表現Hsp27D 的似乎展現了較強的轉移傾向。至於MDA-MB-231 Hsp27 過表現的細胞具有再轉移位點產生細胞聚集的現象並對壓力環 境有較為良好的抵抗性。而在斑馬魚乳癌移植樣本的藥物處理實驗中， 過表現Hsp27 的MDA-MB-231 的組別可以觀察到癌細胞有較強的癌化 傾向。總結來說，Hsp27 的性質再先前的研究中已經用了活體外及老 鼠實驗中證實，而在使用斑馬魚3 天大的乳癌移植實驗中可以看到類 似的結果，說明了斑馬魚對於癌症研究似乎是個可行的模式。
並列摘要
Zebrafish is an animal model which widely utilized in manifold biomedical experiments such as embryo development, hearing loss and toxicity assessment. In recent years, zebrafish has been used for cancer research since the advantage of cost effective and easy caring. Basically, the application of zebrafish in cancer research usually focuses on the property of cancer metastasis and stress resistance ability. Thus, the experiments been conducted would follow this conception mainly. On the other hand, it is been well established that Hsp27 could associate with the ability of enduring the extracellular stresses in the cell, and mainly participate in anti‐apoptosis, cell cycle regulation, and treatment resistance. The activation of downstream mechanism depends on the different type of the Hsp27 multimer conformation (phosphorylation or non‐phosphorylation), which initiate different activity of Hsp27 protein polymer. The aim of the experiment is to attempt to evaluate Hsp27 associated breast cancer activity in the zebrafish model. The 3dpf (day post‐fertilization) zebrafish embryo would be utilize for cancer cell xenograft operation. In order to investigate the Hsp27 phosphorylation pattern affects in breast cancer, the Hsp27D (mimic phosphorylated), and Hsp27A (mimic non‐phosphorylated) had been established, and compare to the result of wtHsp27 (Hsp27 overexpression) and control for further experiments. In BT‐474 breast cancer cell line, expressing Hsp27D seems to have more metastasis tendency than others. As for the MDA‐MB‐231, the result shows that the cell over expressing Hsp27 is inclined to aggregate in metastatic site and shows more resistance upon confronting stresses. Furthermore, zebrafish sample grafting MDA‐MB‐231 with Hsp27 overexpression and MDA‐MB‐231 is also treated with several drugs. The result shows that the group with Hsp27 overexpression remaining more cancerous properties in drug treatment duration compare to control. In sum, several studies had rectified that Hsp27 property in vitro and mice. The result using zebrafish 3dpf embryo for cancer research is similar with previous studies. Illustrating that zebrafish is seems to be a feasible model for cancer research.