2014-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/649385摘要： 儘管在診斷和治療技術上的進步，近年來台灣的癌症盛行率和死亡率還是不斷提高，其主因在於晚期癌症 (advanced cancers) 的腫瘤異質性 (tumor heterogeneity)，造成其對藥物反應不良。更棘手的是，個別腫瘤細胞中，彼此訊息傳遞的合成交互作用 (synthetic interaction) 也不盡相同，因此即使聯合化療及標靶藥物，常常只能殺到部分的腫瘤細胞，成效依然不彰。因此，了解癌症的訊息合成交互作用如何變化，對研發新療法至關重大。 為了解決上述問題，我們將以大腸癌細胞株為平台，研究 FGFR3 的訊息合成交互作用。先前的研究已知纖維母細胞生長因子 (fibroblast growth factor, FGF) 對胚胎發育以及癌症進程相當重要。有趣的是，FGFR3 在不同階段癌症的角色不太相同。首先，在不同期的泌尿道癌症或是大腸直腸癌中，所觀察到的 FGFR3 突變型 (mutant) 或是異形體 (isoforms) 並不一樣。此外，如果在胰臟癌細胞株中大量表現 FGFR3，細胞遷移會取決於其原始型態是像上皮 (epithelial) 細胞還是間質 (mesenchymal) 細胞，來減慢或是加速。因此，我們將用以下方式，探索「FGFR3 在不同情況下有不同功能表現」的分子機轉： 1. 釐清 FGFR3 為何在不同的細胞株有相反的功能。這有兩種可能：不同細胞株可能利用不同的 FGFR3 異形體，驅動不同的訊息傳遞路徑；也有可能不同的 FGFR3 異形體驅動的路徑都相同，但在不同細胞株中與不同的其他路徑交互作用，造成相反的效果。因此，我們將使用多種細胞株，結合多種檢驗方法，包括免疫螢光染色、活細胞影像、RNA interference 等，來找出 FGFR3 對不同細胞有不同影響的秘密。 2. 將同一種細胞作形態轉換，來驗證上述假說。我們將誘導或反轉細胞株的上皮-間質轉形 (epithelial-mesenchymal transition, EMT)，將轉換形態前後的細胞分別拿來檢驗或操弄 FGFR3，以釐清 FGFR3 在不同情況下有不同效果的原因。 3. 檢驗 FGFR3 在不同細胞中是如何調控的。我們將先探討控制 FGFR2 的 ESRP-1和 ESRP-2，對 FGFR3 是否也有影響。此外，我們也將檢驗一些特殊的 FGFR3 異形體和突變型的功能。 透過上述實驗，我們將對細胞的訊息合成交互作用有更深一步的了解，並可進一步據此設計抗癌藥物使用的新策略。 <br> Abstract: The prevalence and mortality of cancer have been steadily increased for decades in Taiwan despite the advance in detection and treatment. It is due to the poor response to treatment for advanced cancers with tumor heterogeneity. Moreover, combination chemo- and targeted therapies have limited benefits due to distinct synthetic interactions between individual tumor cells, resulting in various responses to a uniform treatment protocol. Therefore, understanding how synthetic interactions vary in cancers is critical for the development of novel therapeutics. To solve this problem, we propose to study fibroblast growth factor receptor 3 (FGFR3) using colorectal cancer cell lines. Fibroblast growth factor signaling has been shown important for embryonic development and cancer progression. Interestingly, FGFR3 have distinct roles in various stages of cancer. Distinct isoforms and mutants of FGFR3 have been observed in different stages of urothelial and colorectal cancer. In addition, pancreatic cancer cells over-expressing FGFR3 can slow down or speed up cell migration, depending on whether they are epithelial-like or mesenchymal-like in morphology. We hence aim at unraveling the mechanism how FGFR3 controls cell motility and tumor progression in a context-dependent manner: 1. Clarifying how FGFR3 signaling has distinct effects on the motility of primary human cells and metastatic cancer cells. The distinct roles of FGFR3 may result from different isoforms of FGFR3 having distinct targets to control cell motility. Alternatively, FGFR3 signaling may be all the same in various cell types, but it acts synergistically with other signaling modules distinct among cell types to regulate cell migration differently. We will take multi-disciplinary perturb-and-observe approaches to clarify which hypothesis holds. 2. Validating the above hypothesis by experiments on cells before and after transition. We will induce or reverse epithelial-mesenchymal transition (EMT) in various cell types. In each cell type undergoing transition, its FGFR3 levels & activities will be quantified; its FGFR3 will also be manipulated for its effect on signaling and migration at different stages. 3. Examining the control of FGFR3 signaling and its aberrant forms in various cell types. We will start by examining putative FGFR regulators, such as ESRP-1 and ESRP-2, to see if they also regulate FGFR3. The functional significance of aberrant FGFR3 forms, such as soluble FGFR3 and various FGFR3 mutants will also be examined. Through the above experiments we will have a comprehensive understanding of synthetic interaction in cancer cells, based on which a novel therapeutic strategy can be devised.FGFR3細胞遷移訊息合成交互作用FGFR3cell migrationsynthetic interaction.