PROTEOMIC BIOMARKER PROFILING OF CERVICAL SWABS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 Rader实验室对使用整合的蛋白质组和基因组筛查来寻找宫颈癌的潜在生物标记物感兴趣。宫颈癌筛查非常适合于生物标记物的开发，因为组织采集容易，组织学转换良好。此外，从宫颈样本中获得的细胞和生物液经历了特定的分子变化，这些变化可以被描绘出来，并建立可以预测结果的模型。使用已经在临床实践中使用的标准铲子和细胞刷建立了患者筛查蛋白质和核酸收集方案。使用RNAlater收集样本，然后用蛋白质组学方法鉴定在正常宫颈上皮细胞和宫颈癌细胞中差异表达的生物标志物/蛋白。通过二维差异凝胶电泳法(2-D DGE)鉴定了390个在宫颈癌样本中高水平或低水平(&gt；3倍)表达的斑点。将这些蛋白质组学结果与显微解剖的宫颈肿瘤标本的cDNA微阵列分析中最重要的基因进行比较，以确定重叠的表达模式。最常见的信号通路有：细胞周期：G2/MDNA损伤检查点调控；芳烃受体信号转导；P53信号转导；细胞周期：G1/S检查点调控；以及内质网应激通路。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Rader lab is interested in using integrated proteomic and genomic screening to find potential biomarkers for cervical cancer. Cervical cancer screening is ideally suited for the development of biomarkers due to the ease of tissue acquisition and the well-established histological transitions. Furthermore, cell and biologic fluid obtained from cervix samples undergo specific molecular changes that can be profiled and models constructed that may predict outcome. A patient screening protein and nucleic acid collection protocol was established using standard spatulas and cytobrushes already used in clinical practice. RNAlater was used to collect the samples followed by proteomic methods to identify biomarkers/proteins that were differentially expressed in normal cervical epithelial versus cervical cancer cells. Three hundred ninety spots were identified via two-dimensional difference gel electrophoresis (2-D DIGE) that were expressed at either higher or lower levels (>3-fold) in cervical cancer samples. These proteomic results were compared to the most significant genes in a cDNA microarray analysis of microdissected neoplastic cervical specimens to identify overlapping patterns of expression. The most frequent pathways represented by the combined dataset were: cell cycle: G2/M DNA damage checkpoint regulation; aryl hydrocarbon receptor signaling; p53 signaling; cell cycle: G1/S checkpoint regulation; and the endoplasmic reticulum stress pathway.