实时荧光定量PCR检测大肠癌腹腔微转移及其临床意义_临床医学论文.docVIP

实时荧光定量PCR检测大肠癌腹腔微转移及其临床意义_临床医学论文 实时荧光定量PCR检测大肠癌腹腔微转移及其临床意义_临床医学论文 作者：钱俊 郑树 中西速夫 光幸秀 平井孝 加藤知行【摘要】 目的］ 应用实时荧光定量PCR技术检测大肠癌腹腔内游离癌细胞并探讨其临床意义。［方法］ 2003年5月至2004年1月，收集在日本爱知癌症中心手术81例大肠癌病例的腹腔冲洗液，每个样本的cDNA均应用两种引物合成（随机引物和寡核苷酸引物），并在罗氏实时荧光定量PCR（LightCycler）上进行定量分析。每个样本同时进行常规细胞学检查。所有病例术后经过平均为1年的随访。［结果］ CEA mRNA 和CK-20 mRNA的阳性率和阳性值均与肿瘤的浸润深度（T）、分期以及淋巴结转移相关。在一个合理界定值上，CEA 和CK-20检测的敏感度与特异度均为100%，而常规的细胞学检查则为33%和100%。［结论］ CEA 和CK-20 mRNA定量分析是检测大肠癌腹腔微转移的敏感和特异的方法，CEA 和CK-20 mRNA水平的异常与术后的无瘤生存率显著相关。 【关键词】 大肠肿瘤 癌胚抗原 细胞角蛋白20 实时荧光定量RT-PCR Quantitative Detection of Disseminated Cancer Cells in Peritoneal Washes with Real-Time RT-PCR for Patients with Colorectal Carcinoma and Its Clinical Significance Abstract: ［Purpose］ To detect the free cancer cells in peritoneal cavity in patients with colorectal cancer by quantitative real-time RT-PCR method and investigate its clinical significance. ［Methods］ From May 2003 to Jan. 2004, peritoneal wash was obtained from 81 cases with colorectal cancer in Aichi cancer center. cDNA was synthesized from mRNA extracted from peritoneal washes by 2 kinds of primers (random primer and oligo primer). Quantification of CEA and CK-20 mRNA was performed on a LightCycler instrument with hybridization probe. A part of each peritoneal wash was also examined by conventional cytology. All patients were followed up with an average period of 12 months. ［Results］ Both CEA mRNA and CK-20 mRNA positive rates and values were significantly correlated with depth of tumor invasion, staging, lymph node metastasis. Sensitivity and specificity of CEA and CK-20 real-time RT-PCR with an appropriate cutoff value were all 100%, whereas those of conventional cytology were 33%, and 100%, respectively. ［Conclusion］ Both CEA and CK-20 mRNA quantifications are sensitive and specific methods for detecting micrometastasis in the peritoneal washes with colorectal cancer. Abnormality of CEA and CK-20 mRNA in peritoneal washes is correlated with disease free survival for colorectal cancer. Key