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: 6699653
• 负责人: JAMES D LUKETICH
• 金额: $ 24.22万
• 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/6699653
• 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.

描述（由申请人提供）：食管癌（EC）的发病率是 以前所未有的速度增长。患者预后与淋巴结有关 (LN)参与，但疾病扩散到三个不同的LN领域，使准确的 节点评估困难。传统的成像技术不准确， LN分期。微创LN取样和组织学检查是 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% 复发性疾病的机会。这表明，组织学 检查漏诊了显微镜下的隐匿性转移瘤（CIMs）。本集团与 其他人最近已经表明，使用逆转录病毒的分子检测 转录酶-聚合酶链反应（RT-PCR）与疾病相关 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。我们的初步数据表明，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LN阴性患者， 复发的风险。我们建议使用QRT-PCR来检测我们的大型 EC患者群体，并将QRT-PCR与常规组织学进行比较。在 具体目标1我们将测量分子标记物的mRNA表达水平 癌胚抗原（CEA）和细胞角蛋白19（CK 19）在LN的EC和 良性疾病患者。有了这些信息，我们将确定最佳的 用于定义阳性LN的表达水平截止值。然后我们将 比较两种方法预测复发的敏感性和特异性 LN的QRT-PCR和组织学分析。组织学诊断为 N1疾病提供辅助放化疗。高达50%的人可能有一个 病理完全缓解，但其中超过50%会复发。在 具体目的2、确定QRT-PCR的临床意义 在接受过化疗的组织学分期降低的患者中， N1 EC的新辅助治疗。最后，术中LN分期 冷冻切片的组织学检查不能检测到病毒，错误率为7- 10%。 与固定的组织学相比，在第三阶段，我们将 探索QRT-PCR技术（SmartCycler）的新发展， 提供LN的快速术中QRT-PCR分析，并提高准确性 手术分期本研究将确定QRT-PCR在以下方面的作用： 对EC患者的LN进行分期，确定临床复发是否可以 在新辅助治疗病理分期降低的患者中预测 评价术中QRT-PCR的结果及临床应用价值。