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

描述(由申请人提供)：这项提案的主要目标是确定microRNAs是否参与肺癌药物敏感性相关关键基因的调节。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，我们预测一些癌基因、肿瘤抑制基因和与耐药相关的基因都受到miRNAs的调控。我们已经使用miRNA在少数细胞系上的表达谱数据证实了其中几个预测，包括小细胞肺癌(SCLC)、非小细胞肺癌(NSCLC)和永生化的人支气管上皮(HBEC)细胞系。我们将使用计算机和体外相结合的方法来解决以下问题：肺癌细胞系中特定的microRNA表达谱是否与不同的药物敏感性相关？MicroRNAs在肺癌细胞的药物敏感性/耐药性中起作用吗？能否通过操纵microRNA的表达水平来提高细胞培养中肺癌细胞的药物敏感性？研究设计：我们将使用microRNA微阵列来识别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控的miRNAs的mRNA靶标。我们将使用报告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目标mRNA的抑制翻译有关。最后，我们将确定microRNAs的过度表达或沉默是否会影响目标基因(S)的mR NA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：识别与miRNA参与癌症的模型一致的异常miRNA表达可以为肺癌疾病机制提供新的见解，并提供一套新的诊断标志物和治疗靶点，可能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.