microRNA regulation of drug sensitivity in non-small cell lung cancer

描述（由申请人提供）：本提案的主要目的是确定microRNA是否参与调节与肺癌药物敏感性有关的关键基因。我们已经开发了一种计算方法来预测miRNAs和mRNAs之间的相互作用。基于这种方法，我们预测了一些癌基因，肿瘤抑制基因，以及与耐药相关的基因受到miRNA的调控。我们已经证实了这些预测中的几个，使用小细胞肺癌（SCLC），非小细胞肺癌（NSCLC）和永生化人支气管上皮（HBEC）细胞系的小RNA表达谱数据。我们将使用计算机模拟和体外方法的组合来解决以下问题：特异性microRNA表达谱与肺癌细胞系的差异药物敏感性相关吗？microRNA在肺癌细胞的药物敏感性/耐药性中发挥作用吗？microRNA表达水平可以被操纵以增加细胞培养中肺癌细胞的药物敏感性吗？研究设计：我们将使用microRNA微阵列来鉴定在一组肺癌细胞系中差异表达的miRNAs。然后，我们将使用我们自己的生物信息学工具以及其他研究人员开发的工具来预测受调节的miRNA的mRNA靶点。我们将使用报告基因分析系统在培养细胞中进行研究，以确定miRNA的表达是否与培养细胞中预测的靶mRNA的抑制翻译相关。最后，我们将确定microRNA的过度表达或沉默是否会影响靶基因的mRNA翻译，并导致对广泛使用的抗癌药物敏感性的特定变化。癌症相关性：鉴定与miRNA参与癌症的模型一致的异常miRNA表达可以提供对肺癌疾病机制的新见解，以及一组新的诊断标记物和治疗靶点，其可以在人类疾病的治疗中发挥重要作用。

DESCRIPTION (provided by applicant): The primary objective of this proposal is to determine if microRNAs participate in the regulation of critical genes implicated in lung cancer drug sensitivity. We have developed a computational method to predict interactions between miRNAs and mRNAs. Based on this method, we predict that a number of oncogenes, tumor suppressor genes, and genes relevant to drug resistance are regulated by miRNAs. We have confirmed several of these predictions using miRNA expression profiling data on a small number of cell lines, including small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC) and immortalized human bronchial epithelial (HBEC) cell lines. We will use a combination of in silico and in vitro approaches to address the following questions: Are specific microRNA expression profiles correlated with differential drug sensitivity in lung cancer cell lines? Do microRNAs play a functional role in drug sensitivity/resistance of lung cancer cells? Can microRNA expression levels be manipulated to increase drug sensitivity of lung cancer cells in cell culture? Study design: We will use microRNA microarray to identify miRNAs that are differentially expressed across a panel of lung cancer cell lines. We will then predict the mRNA targets of the regulated miRNAs using both our own bioinformatics tools as well as those developed by other investigators. We will perform studies in cultured cells using a reporter assay system to determine if expression of the miRNA is associated with repressed translation of the predicted target mRNA in cultured cells. Finally, we will determine if over-expression or silencing of the microRNAs affects mRNA translation of the target gene(s) and results in specific changes in sensitivity to widely-used anti-cancer drugs. Cancer relevance: Identifying aberrant miRNA expression consistent with the models of miRNA involvement in cancer can provide new insight into lung cancer disease mechanisms, and a new set of diagnostic markers and therapeutic targets which could play a significant role in the treatment of human disease.