Use of Quantitative RT PCR for Staging Esophageal Cancer

使用定量 RT PCR 对食管癌进行分期

• 批准号: 6514992
• 负责人: JAMES D LUKETICH
• 金额: $ 27.08万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-09 至 2005-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6514992
• 关键词: biopsy; carcinoembryonal antigen; clinical research; esophagus neoplasm; genetic markers; human subject; lymph node neoplasm; metastasis; neoplasm /cancer classification /staging; neoplasm /cancer genetics; polymerase chain reaction

DESCRIPTION (Provided by applicant): The incidence of esophageal cancer (EC) is rising at an unprecedented rate. Patient prognosis is related to lymph node (LN) involvement, but disease spread to three distinct LN fields makes accurate nodal assessment difficult. Conventional imaging techniques are inaccurate for LN staging. Minimally invasive LN sampling with histological examination is currently the most accurate staging protocol, but even with radical LN dissection, patients with histologically negative LN have a 30-50 percent chance of developing recurrent disease. This suggests that histologic examination misses microscopically occult metastases (MOM). Our group and others have recently shown that molecular detection of MOM using reverse transcriptase-polymerase chain reaction (RT-PCR) correlates with disease recurrence, but the clinical relevance of these findings is debatable due to a high false positive rate. Our preliminary data shows that quantitative RT-PCR (QRT-PCR) can discriminate between true, and false positives, and thus can accurately identify histologically LN negative patients who have an increased risk of recurrence. We propose to use QRT-PCR to detect MOM in our large population of EC patients, and compare QRT-PCR with conventional histology. In Specific Aim 1 we will measure mRNA expression levels of molecular markers carcinoembryonic antigen (CEA) and cytokeratin 19(CK19) in LN's from EC and benign disease patients. With this information, we will determine the optimal expression level cut-off values for defining positive LN's. We will then compare sensitivity and specificity for recurrence prediction using both QRT-PCR and histologic analysis of LN's. Patients diagnosed with histologically N1 disease are offered adjuvant chemoradiotherapy. Up to 50 percent may have a pathologic complete response but of these, over 50 percent will recur. In Specific Aim 2 we will determine the clinical significance of QRT-PCR positivity in LN's of histologically downstaged patients who have received neoadjuvant treatment for N1 EC. Finally, intraoperative LN staging by histology of frozen sections does not detect MOM, and is incorrect 7-10 percent of the time compared with fixed tissue histology. In Specific Aim 3 we will explore the use of new developments in QRT-PCR technology (SmartCycler), to provide rapid, intraoperative QRT-PCR analysis of LN's and improve the accuracy of surgical staging. This investigation will define the role of QRT-PCR in staging LN's from patients with EC, determine if clinical recurrences can be predicted in patients who were pathologically downstaged by neoadjuvant therapy and assess results and clinical applications of intraoperative QRT-PCR.

资料(申请人提供)：食道癌发病率为 以前所未有的速度上升。患者的预后与淋巴结有关 (LN)参与，但疾病传播到三个不同的LN领域使准确 节点评估困难。传统的成像技术不能准确地 在舞台上。微创取材结合组织学检查是 目前最准确的分期方案，但即使是激进的LN 解剖，组织学阴性的LN患者有30%-50% 罹患复发性疾病的几率。这表明组织学上 检查漏掉了显微镜下的隐匿性转移瘤(MOM)。我们的团队和 其他人最近表明，使用反向检测MOM的分子 逆转录聚合酶链式反应(RT-PCR)与疾病 复发，但这些发现的临床相关性是有争议的，因为 假阳性率高。我们的初步数据显示，定量RT-PCR (qRT-PCR)可以区分真阳性和假阳性，因此可以 准确识别组织学上LN阴性患者 复发的风险。我们建议使用QRT-PCR法来检测大鼠的MOM 并与常规组织学方法进行比较。在……里面 具体目标1我们将测量分子标记的mRNA表达水平 癌胚抗原(CEA)和细胞角蛋白19(CK19)在EC和LN中的表达 良性疾病患者。有了这些信息，我们将确定最佳的 定义正LN的表达式级别分界值。然后我们将 比较两种方法预测复发的敏感性和特异性 组织学诊断为狼疮性肾炎患者的QRT-PCR及组织学分析 为N1疾病提供辅助化疗和放射治疗。高达50%的人可能会有 病理完全缓解，但其中超过50%会复发。在……里面 具体目的2我们将确定QRT-PCR的临床意义 组织学分期较低的患者接受手术后LN的阳性表达 N1EC的新辅助治疗最后，术中LN分期通过 冰冻切片的组织学检测不到MOM，不正确的是7%-10% 与固定组织学相比较的时间。在具体目标3中，我们将 探索QRT-PCR技术(SmartCycler)的新发展，以 提供快速、术中LN的QRT-PCR分析，并提高准确性 手术分期的结果。这项研究将确定QRT-PCR在 对EC患者的LN进行分期，确定临床复发是否 新辅助治疗病理分期降低的患者的预测 评价术中QRT-PCR的结果及临床应用。