2013-10-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/678390摘要：癌症不僅是台灣首位死亡原因也是世界的大問題。許多病患初次診斷癌症時就已合併癌細胞轉移，癌細胞轉移通常也導致較差的疾病預後，癌轉移始於癌細胞侵襲周邊組織，癌細胞侵襲發生在非常早期，與癌細胞及微環境間的互動有密切關聯。微環境主要由間質細胞與細胞外間質構成，細胞外間質是由大分子物質形成的複雜網狀結構，具有特定生物物理化學及生物力學特性，細胞外間質調節異常對癌症進展與轉移具有關鍵性角色。從臨床的角度，儘早抑制癌細胞侵襲，限制癌症進展與癌細胞轉移，可以改善癌症治療結果。但是由於缺少適合的研究工具模型，使我們難以深入瞭解癌細胞侵襲與細胞外間質調節異常的全貌，而限制了治療的進展。 我們嘗試建立三維細胞培養影像術，並量化測量癌細胞侵襲及細胞-細胞外間質互動。光聲分子影像之基本原理是能量轉換的物理原則，分子將光能吸收轉換為聲波，進一步利用超音波收集物質分子震動聲波轉為影像。配合奈米金粒子在光聲分子影像中可產生特定訊號之特性，以及三維細胞培養架構模擬細胞-細胞外間質互動，光聲分子影像就能重建癌細胞侵襲與細胞外間質調節異常影像。超音波剪力波彈性藉由細胞外間質彈性變化，可測量細胞外間質物理及生化性質的變化。利用影像重組技術，光聲分子及超音波剪力波彈性變化影像能夠量化測量，並依時間性及空間性分析三維細胞培養架構中(1)腫瘤細胞、間質細胞位置變化(2)胞外間質之型變度(3)剪力波彈性變化比對圖及(4)壓力場之變度。 目前常用評估癌細胞侵襲的工具，多著重於細胞遷移而未觸及細胞外間質變化與細胞-細胞外間質互動。合併三維細胞培養系統模擬細胞-細胞外間質互動、光聲分子影像及剪力波彈性影像，不僅追蹤細胞遷移與細胞外間質調節異常，也可深入探討癌細胞侵襲與細胞外間質之交互作用。本計畫主要目的在(1)建立三維細胞培養系統模擬癌細胞-細胞外間質互動(2)尋找調控細胞-細胞外間質互動之物理及生化因子(3)以三維細胞培養系統模擬癌細胞-細胞外間質互動探索癌症治療能力。如此有助於我們釐清癌細胞在調節異常的細胞外間質中之進展狀況，以及在癌細胞侵襲時細胞外間質的相對應變化。進一步我們可以確認腫瘤化學預防藥物以及藥物治療模式之效用，並加速化學預防藥物之新藥篩選開發，也有機會發展個人化抗腫瘤藥物治療。<br> Abstract: Cancer disease is not only the leading cause of deaths in Taiwan but also worldwide. A numerous of patients with newly diagnosed solid cancer already have clinically detectable metastases. Metastasis is initiated by invasion of primary cancer cells into the adjacent tissue and leads to a poor disease prognosis. Cancer cell invasion may take place in very early stage of cancer formation and is closed related interaction between cancer cell and their microenvironment. The major components of microenvironment are various stromal cells and extracellular matrix (ECM), which is a complex network of macromolecules with distinctive biophysical, biochemical and biomechanical properties. The deregulated ECM plays a central role in cancer progression and metastasis. In the clinical setting, inhibition of cancer cell invasion as early as possible will improve the outcome of malignant diseases by limiting cancer expansion and reducing cancer metastases to distant sites. But poor understanding about cancer invasion and ECM deregulation may result from lacking of proper tool or model for investigation and limit the improvement of disease treatment. We try to establish a 3D cell culture system imaging and intend to measure cancer invasion and cell-ECM interaction quantitatively. The photoacoustic molecular imaging (PAMI) is based on energy transfer—energy of light can be absorbed by molecules and the energy transfer as acoustic signal collected by vector and reconstructed as images. Combination with gold nano-rods (AuNRs), that show specific signals in PAMI, and development of 3D cell culture system that mimic cell-ECM interaction, the cancer invasion and ECM deregulation can be reconstructed by PAMI. The shear wave elasticity imaging (SWEI) can measure the physical or biochemical changes of ECM through elasticity (stiffness) alteration. Collection of snap image and image reconstruction, PAMI and SWEI offer quantitative measurement about (1) cancer and stromal cells location, (2) deformation field, (3) shear modus mapping and (4) stress field in 3D cell culture scaffolds spatially and temporally. Now existing assay tools for cancer invasion, almost evaluate the cancer cell migration but not alteration of ECM and cell-ECM interaction. Combination of 3D culture system mimic cell-ECM interaction, PAMI and SWEI, not only tracking cells migration but also evaluating ECM deregulation, we can explore cancer invasion and cell-ECM interaction deeply. The implementation of this project comprises three main aims: i) Building in vitro 3D cell culture system that mimics cancer cell-ECM interactions in tumor milieu. ii) Searching for physical and biochemical factors that regulate in vitro 3D cancer cell-ECM interactions. iii) Exploring the therapeutic potential of in vitro 3D cancer cell-ECM interaction model. These will help us clarifying cancer invasion in a deregulated ECM and the reciprocal alteration of ECM before, during and after cancer cell invasion. Furthermore, we can verify the therapeutic potential of chemoprevention agents, therapeutic strategies and accelerating drug screen of chemoprevention agent candidates. And those provide the opportunity to develop personalized medication on cancer therapy.癌細胞侵襲細胞-細胞外間質互動三維細胞培養系統光聲分子影像剪 力波彈性影像Cancer invasionCell-ECM interaction3D cell culture systemPhotoacoustic molecular imaging (PAMI) and Shear wave elasticity imaging (SWEI)