2011-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/660015摘要：癌症在世界上或台灣皆為造成人類死亡的第一位，無論癌症診斷與治療的進步，患者的治療成果與存活率仍令人失望。近年來，分子標靶療法提供了對抗癌症的機會，但是，要發現如肺癌的EGFR 和血癌的BCR-ABL tyrosine kinase 等好的分子標靶標的基因，可以針對某些癌症有相當好的療效，有一定的難度。因此，如Atlas 等大型計畫運用最新的技術-新一代定序儀分析數種癌症的基因序列，希望可以研發新的分子標靶標的基因。不幸地，這些耗資不斐的計畫並沒有達成當初設定的目標。本計畫的目標是發展出一個據成本效益的研究方法。藉此，我們可以發展出癌症的預後印記。微陣列技術的出現為二十世紀的生物科學與生物醫學帶來極大的衝擊，它可以高通量地偵測基因表現、基因拷貝數變異、表觀基因修飾與微核醣核酸表現，而這些由微陣列技術所衍生的眾多資料所代表的生物醫學意義，有賴快速發展的生物資訊與予分析，我們可以利用這些分析資料研發疾病預後印記與新藥開發。先前，我們運用微陣列與一系列具有不同侵襲能力的肺癌細胞株，發現了數十個與肺癌轉移相關的基因。NCI-60 細胞株包含了59 種人類的各式的癌症細胞株，為體外抗癌藥物篩選最廣為使用的細胞株，同時，這些細胞株的基因表現、基因拷貝數變異、表觀基因修飾與微核醣核酸表現業經由各式的方法所測量而公布於公開網站。有3 種細胞特徵會左右癌症的惡性度，分別為細胞侵襲性、血管新生和腫瘤聚落形成。高涵量篩選系統是一種以細胞為基礎的嶄新篩選工具，它可以同時間快速地擷取並分析多孔盤中細胞的多重參數。在本研究中，我們首先將建立以高涵量篩選系統分析細胞侵襲性、血管新生和腫瘤聚落形成的平台，以及建立啟動子甲基化的微陣列平台，之後，我們將結合細胞性狀的資料與網路免費下載的NCI-60 的微陣列資料，分析出與細胞侵襲性、血管新生和腫瘤聚落形成相關的基因體、表觀基因體和轉譯體上的變異，我們進一步評估這些基因或微核醣核酸對癌症惡性度的影響，並分析他們的可能調控機制。最後，將利用這些與癌症性狀相關的基因表現、基因拷貝數變異、表觀基因修飾與微核醣核酸表現發展為癌症病患預後印記。這些所有的努力將有助於我們解開癌症的分子機制與發展癌症的治療策略。<br> Abstract: Cancer is the leading cause of human death in Taiwan and worldwide. Despite of advancesin the diagnosis and therapy of cancers, the therapeutic outcome and patients’ survival arefrustrated. Recently, molecular targeting therapy (MTT) provided an opportunity to fightagainst cancers but it is difficult to identify the good MTT targets like EGFR in lung cancerand BCR-ABL tyrosine kinase in leukemia against a major fraction of the subset cancers.Hence, several large projects (such as Atlas) use the most advance technique, NextGeneration Sequencer, to analyze genomic sequences of several cancers. Unfortunately,these projects do not meet the original goals in spite of the huge budget funded. The goal ofthis project is to develop a cost-effective approach by which we can develop the prognosticsignatures for cancers. Microarray technology has made tremendous impact in biologicalscience and medicine in the 2000’s. Microarray can allow for high throughput determinationof gene expression, gene copy number variation, epigenetic modification and microRNAexpression. The biomedical relevance of huge amount of data from microarray studies isfurther facilitated by the vigorous growth of bioinformatics. We can use this information toidentify signatures for prognosis prediction and new drug targets. Previously, we usemicroarrays and a series of lung cancer cells with varying invasion ability to identify dozensof metastasis-related genes. The NCI-60 cell line set including 59 diverse human cancer celllines is one of the best characterized cell line sets available for anticancer compoundscreening which has been widely used for in vitro anticancer drug testing. These also havebeen analyzed by using a variety of methods including transcriptional profiling, epigeneticDNA methylation, gene copy number variation and microRNA profiling. There are 3features important to cancer malignancy, tumorigenesis, angiogenesis and metastasis. TheHigh Content Screening System (HCS) is an advance cell-based screening tool which is usedto rapidly acquire and analyze the cell images in multiple parameters in multiple well plates.In this project, we will establish in vitro invasion, angiogenesis and colonogenesis assays byHCS system and setup the promoter methylation microarray system. we then integratecancer phenotyping data and the microarray data of NCI-60 free downloaded to identify thegenomic, epigenomic and transcriptomic aberrations related to cancerous phenotypesincluding invasion, colony formation and tube formation capabilities, further to evaluate theeffect of identified genes/microRNAs on cancer malignancies, to elucidate the underlyingregulatory mechanisms and to develop the prognostic signatures for clinical prediction ofcancer patients’ outcome. All these efforts will help us to interpret the molecular mechanismsof cancers and develop therapeutic strategy of cancers.高涵量篩選系統癌侵襲血管新生癌轉移癌症預後印記High Content Screening Systeminvasionangiogenesistumorigenesiscancer prognostic signature