CHARACTERIZATION OF DICER MRNA VARIANTS IN ORAL CANCER

口腔癌 DICER mRNA 变异的特征

• 批准号: 8360488
• 负责人: Andrew George Jakymiw
• 金额: $ 17.58万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-09-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360488
• 关键词: Address; Apoptosis; Biological; Biological Assay; Cancer Biology; Cell Line; Cell Proliferation; Cells; Centers of Research Excellence; Code; Data; Development; Disease Progression; Enzymes; Eukaryota; Funding; Genes; Grant; Human; Lead; Malignant Epithelial Cell; Messenger RNA; Metric; MicroRNAs; National Center for Research Resources; Oncogenic; Oral; Oral health; Pathway interactions; Pattern; Play; Principal Investigator; Property; RNA Interference; RNA Splicing; Relative (related person); Reporting; Research; Research Infrastructure; Resources; Ribonuclease III; Role; Source; South Carolina; Tissues; United States National Institutes of Health; Variant; cancer cell; cancer type; cell motility; cost; human DICER1 protein; insight; keratinocyte; mRNA Expression; malignant mouth neoplasm; mouth squamous cell carcinoma; overexpression; protein expression

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Dicer is a highly conserved RNase III type enzyme found in almost all eukaryotes that is essential for the RNA interference (RNAi) and microRNA pathways. During the last several years an increasing number of reports have found Dicer to be aberrantly expressed in different types of cancer, including oral squamous cell carcinomas (OSCCs). Recently, the dicer gene has been predicted to produce 11 mRNA splice variants bearing modified coding sequences. Our preliminary data suggests that one of these predicted Dicer mRNA splice variants is expressed in cells and appears to be upregulated in OSCC cell lines. Because the expression and function of the Dicer mRNA splice variants have not been well characterized and it currently remains unclear as to their biological significance, this proposal will explore the role of this particular Dicer mRNA splice variant in relation to oral cancer biology. Therefore, to better assess and correlate the expression patterns of the Dicer mRNA splice variant relative to OSCCs and disease progression this proposal will characterize its mRNA and protein expression levels in both OSCC cell lines and tissues ranging from dysplastic to invasive OSCCs in relation to non-cancerous counterparts. Furthermore, to enhance our understanding of the mechanisms regulating its aberrant expression this study will also analyze whether its aberrant expression in oral cancer cells is due to transcriptional and/or post-transcriptional regulatory mechanisms. Because the significance of the dysregulation of this splice variant in terms of cancer cell properties is not understood, this proposal will further seek to examine the biological effects of reducing the levels of the Dicer mRNA splice variant in oral cancer cells using RNAi knockdown strategies and analyze the effects of overexpressing it in both primary and immortalized human oral keratinocytes. The biological metrics will include cell proliferation, apoptosis, cell migration and invasion, and oncogenic transformation assays. By successfully addressing these specific aims this will lead to a better understanding of oral cancer biology. Furthermore, it will shed new insights into the function of Dicer mRNA splice variants and what roles they play in oral cancer development and progression.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 DICER是一种在几乎所有真核生物中发现的高度保守的RNase III型酶，这对于RNA干扰（RNAI）和microRNA途径至关重要。 在过去的几年中，越来越多的报告发现DICER在不同类型的癌症中异常表达，包括口服鳞状细胞癌（OSCC）。 最近，已经预测DICER基因会产生11个带有改良的编码序列的mRNA剪接变体。 我们的初步数据表明，这些预测的DICER mRNA剪接变体之一在细胞中表达，并且似乎在OSCC细胞系中上调。 由于迪切尔mRNA剪接变体的表达和功能尚未得到很好的特征，目前尚不清楚它们的生物学意义，因此该建议将探讨该特定的DICER mRNA剪接变体在与口腔癌生物学方面的作用。 因此，为了更好地评估和关联DICER mRNA剪接变体的表达模式相对于OSCC和疾病的进展，该提案将表征其在OSCC细胞系和从异型增生到侵入性OSCC的组织中其mRNA和蛋白质表达水平相对于非癌性对抗。 此外，为了增强我们对调节其异常表达的机制的理解，本研究还将分析其在口腔癌细胞中的异常表达是否是由于转录和/或转录后调节机制引起的。 由于尚不理解这种剪接变体在癌细胞特性方面的失调的重要性，因此该提案将进一步寻求使用RNAi敲低策略中降低口腔癌细胞中迪切尔mRNA剪接变体水平的生物学效应，并分析其在原发性和永生化的人类口腔氨基甲酸中含量过表达的影响。 生物指标将包括细胞增殖，凋亡，细胞迁移和侵袭以及致癌转化测定法。 通过成功解决这些特定目标，这将使人们对口腔癌生物学有更好的了解。 此外，它将为DICER mRNA剪接变体的功能以及它们在口腔癌发展和进展中起什么作用提供新的见解。