Regulation of mRNA Stability in Human Saliva

研究正常唾液与癌症唾液中 mRNA 的表达和调控，为发现口腔癌诊断标记物带来了巨大潜力。毫无疑问，唾液是最容易收集和保存的非侵入性体液——之前的研究表明，唾液中含有人类mRNA，可用于口腔癌的诊断生物标志物。该技术基于微阵列，非常适合口腔病理学的实时医学诊断。该提案的目标是了解人类唾液中 mRNA 稳定性和不稳定的作用。我们通过微阵列鉴定的大多数口腔癌基因在其 3' UTR 序列中含有富含 AU 的元件 (ARE)。 ARE 靶向 mRNA，通过其反式作用蛋白快速降解。因此，该提案的主要目的是鉴定唾液中的 mRNA 结合蛋白以及该过程涉及哪些因素。在癌症条件下，唾液中含有 ARE 的转录物上调并稳定。因此，我们的假设是，ARE 转录本通过避免 mRNA 衰减机制在癌症条件下保持稳定。我们计划利用 mRNA 降解系统以及我们之前对 mRNA 降解途径的研究中获得的专业知识来进一步测试、验证和识别唾液中 mRNA 稳定性的机制。该提案包括以下目标： I) 阐明与人类唾液中 mRNA 稳定性有关的因素。 II) 确定癌症患者唾液中 mRNA 稳定的基本机制，III) 验证包含 mRNA 衰减的 ARE 与 MAP 激酶/通路激活的相关性。总之，这些研究将揭示正常和口腔癌中 mRNA 的稳定性，从而推动癌症 RNA 生物标志物的临床诊断取得进展。

Studying expression and regulation of mRNA in normal versus cancer saliva is garnering great potential for the discovery of diagnostic marker for oral cancer. Undoubtedly, saliva is the most non-invasive body fluid that can be easily collected and preserved- Previous research has shown that saliva contains human mRNAs that can be used for diagnostic biomarkers for oral cancer. This technology is a microarray based and well suited for real-time medical diagnostics for oral pathology. The goal of this proposal is directed toward understanding the role of mRNA stability and de-stability in human saliva..The majority of oral cancer gene's which we identified through microarray contain AU-rich elements (AREs) at their 3' UTR sequences. AREs target mRNAs for rapid degradation via its trans-acting proteins. Hence the main aim of this proposal is to identify mRNA binding proteins in saliva and what factors are involved in this process. Under cancer conditions ARE containing transcripts are up-regulated and stabilized in saliva. Thus, our hypothesis is that ARE transcripts are stabilized in cancer conditions by means of avoiding mRNA decay machinery. We plan to utilize mRNA decay system along with expertise gained from our previous studies on mRNA degradation pathways so far to further test, validate and identify the mechanistic aspects of mRNA stability in saliva. The proposal includes the following aim: I) Elucidate the factors involved in mRNA stability in human saliva. II) Determine the fundamental mechanism responsible for the stabilization of mRNA's in saliva of cancer patients, III) Validate the ARE containing mRNA decay in correlation with MAP kinase/pathway activation. In summary, these studies will open up the mRNA stability in normal and oral cancer lead to advances in clinical diagnostics of RNA biomarkers in cancer.