2011-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/657848摘要：肺癌在台灣各種癌症的致死率中高居第一位。雖然肺癌診斷及治療不斷的有新進展，肺癌的發生率及死亡率仍逐年增加。造成肺癌高死亡率的主因是肺癌診斷時往往已經轉移，大約只有15%的肺癌病患有機會接受手術治療。而有機會接受手術治療的病患也只有一半可以長期存活，其餘病患往往死於癌症轉移。如何早期診斷肺癌且給予有效的治療，並找出轉移率高的危險群施以有效的輔助性治療，將有助於減少肺癌的死亡率。目前對於癌症的型成與轉移，有越來越多的證據顯示與癌細胞周圍組織以及間質細胞間之交互作用有密切關係。其中以腫瘤相關纖維母細胞由於與癌症的型成與轉移有關而受到較多之關注。為了研究癌細胞與纖維母細胞之交互作用，吾人利用CL1-0與HS 68兩種細胞株同時培養並觀察。於是吾人發現HS 68細胞會被CL1-0細胞活化，同時CL1-0細胞也伸出偽足爬向HS 68細胞並且互相接觸。在接觸的同時發現接觸的周圍有微泡(microvesicles)的產生。另外利用microfluidic cell culture chip觀察CL1-0與MRC-5細胞之交互作用，發現CL1-0之培養液會刺激MRC-5持續性活化。而CL1-0細胞只能被活化的MRC-5培養液刺激發生非持續性之移動性增加。由於以上的發現，吾人希望進一步利用微陣列及代謝體技術探討造成纖維母細胞活化以及肺癌細胞爬行之因子。另外也將利用質譜儀分析微泡內可能之成分，進而追蹤微泡型成以及運輸傳送之過程。希望藉由對於癌細胞與腫瘤相關纖維母細胞間互動之了解，找出可用來治療肺癌的新標的。<br> Abstract: Lung cancer is the leading cause of cancer deaths worldwide. Despite the fact that early stage disease may be cured with surgery, Only 15%-20% of lung cancer patients were operable at presentation and about half of them had long term survival. The remaining almost died of metastatic disease, even though the initial pathologic staging is favorable. There is growing evidence that carcinogenesis and metastasis are controlled by cellular interactions derived from a complex relationship between stromal, epithelial, and extracellular matrix (ECM) components. Among these different cell-types, cancer-associated fibroblasts (CAFs) are attracting increasing attention both as recipients and as producers of pro-tumorigenic signals. Activated fibroblasts in the stroma promote tumor progression by secreting growth factors and pro-migratory ECM components, as well as upregulating the expression of serine proteases and matrix metalloproteinases that degrade and remodel the ECM. To investigate the interaction of CAFs with lung cancer cells, we conducted human lung adenocarcinoma cell line CL1-0 and human fibroblast cell line HS 68 and co-culture experiments. Our preliminary data showed 1) HS68 cells were activated by CL1-0 cells, 2) CL1-0 cells increased motility after co-culture, 3) CL1-0 cells contacted with HS 68 cells directly after co-culture with microviscles transportations between cancer cells and activated fibroblasts. We conducted microfluidic cell culture chip experiment to investigate the timing sequences of paracrine loop between lung cancer cells ad fibrobla. We found that lung cancer cells activate the fibroblasts, and then the activated fibroblasts enhance the migration speeds of cancer cells by paracrine stimulation. Once the fibroblasts are activated, this phenotype can persist without sustained stimulation by cancer cells. However, the high-speed migration of cancer cells cannot continue without the paracrine stimulation of activated fibroblasts. The current proposal aims to explore the paracrine signaling pathways between lung cancer cells and fibroblasts, as well as the mechanism of maintenance of fibroblast activation by way of Oligonucleotide microarray and metabolomics. For direct cell contact and microvesicles transportation, we will isolate microvesicles and determined the protein/peptide contents by LC-MS/MS. Further tracking for microvesicle transportation will be conducted by fluorescence tagged factors within microvesicles. With the above approach, we aim to explore the paracrine signaling and mechanism of direct contact and microvesicle transportation between cancer cells and CAFs. With the knowledge of how CAFs are recruited in cancer microenvironment and how CAFs support tumorigenesis and metastasis, we could develop novel target in lung cancer treatment.肺癌腫瘤相關纖維母細胞腫瘤相關巨噬細胞機械性牽張代謝體學轉移lung cancercancer-associated fibroblasttumour-associated macrophagemechanical stretchingmetabolomicsmetastasis