2019-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/699094摘要：利用低劑量電腦斷層做為肺癌篩檢可以降低20%的死亡率。但是偽陽性高達 96.4%，也就是說大部分電腦斷層看到的異常影像絕大部分為良性。增加肺癌篩檢的敏感性與特異性是臨床醫師仍未完成的夢想。利用非侵襲性或侵襲性較小的方法所得的特殊標記做為癌症診斷，可能是解決的方法之一。基因啟動子(promoter)的甲基化可以抑制基因的表現，進而影響肺癌的形成。之前對於肺癌基因甲基化的研究，主要來自臨床檢體整塊肺癌組織或是肺癌細胞株。這樣的研究常忽略了癌細胞周邊微環境的影響，特別是癌相關纖維母細胞。在我們之前的研究，建立了許多癌相關纖維母細胞（CAF）及成對的正常纖維母細胞（NF）。全面性的DNA甲基化分析顯示與NF相比，CAF中有數百個重要基因高甲基化或低甲基化。氣管灌洗液及吐氣濃縮液臨床上還是容易收集的檢體。這兩種檢體皆含有氣道細胞外的液體（extracellular lining fluid），可能直接來自於癌細胞或癌細胞周遭的微環境。在本研究，我們將收集BAL及EBC。特殊基因的甲基化，包括抑癌基因，或是CAF中、高甲基化的基因將被測試。首先必須先確定所收集的檢體確實含有微量DNA，其次確定P16的甲基化做為positive control。接著分析有興趣基因的甲基化。期望藉由本研究找到DNA甲基化標記，做為肺癌早期偵測及預測預後的工具。<br> Abstract: Lung cancer screening by low dose computer tomography can reduce 20% mortality. However, the false positive rate is up to 96.4% that mean most of the positive findings are benign lesions. Increase the sensitivity and specificity is unmet need in clinical practice. Biomarker from non-invasive or less invasive methods is one of solution. Methylation in gene promotor causes stable silencing of associated genes and is an important epigenetic event in the development of lung cancer. Previous studies examining whole lung cancer tissue or cancer cell lines have omitted the microenvironment especially cancer associated fibroblast (CAF). In our previous study, we had established a lot of CAFs and paired normal fibroblast (NF). Comprehensive DNA methylation profiling showed hundreds of significant genes that were hyper-methylated or hypo-methylated in CAFs when compared to NFs. Bronchial alveolar lavage (BAL) and exhaled breath condensate (EBC) are easy to collect. BAL and EBC both contain extracellular lining fluid possible comes from cancer cells and cancer microenvironment. In this study, we want to collect the specimens from respiratory tract via non-invasive (EBC) or less invasive methods (BAL). Methylation of specific genes including the oncosuppressor and the genes which were hyper-methylated in CAFs will be checked in collected samples. First step will be DNA detection in minutes samples, followed by p16 methylation detection. Finally, we will test all the methylation of candidate genes. We hope to find DNA methylation markers for early detection of lung cancer and for prognosis prediction.甲基化生物標記肺癌氣管灌洗液吐氣濃縮液methylationbiomarkerlung cancerbronchial alveolar lavageexhaled breath condensate