Detection and clinical application of circulating cancer cells, using highy pricise cancer cell determination system

循环癌细胞的检测及临床应用，采用高性价比的癌细胞测定系统

基本信息

批准号：
13470241
负责人：
MAEHARA Yoshihiko
金额：
$ 9.28万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470241/
关键词：
Real-time PCR CEA CK18 CK19 CK20 Bone marrow Peripheral blood 標的遺伝子サイトケラチン18 定量的RT-PCR 臨床病理学的解析リアルタイムPT-RCT 微小転移

项目摘要

Real time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect disseminated cancer cells in bone marrow and peripheral blood. First of all, we compared CEA, CK18 and CK20 expression using 4 gastric cancer cell lines and 3 normal tissue cell lines in order to select the most appropriate marker for detection of disseminated gastric cancer cell in bone marrow. CK20 proved to be the most promising marker since the expression level of normal cell lines was extremely low and about 50- to 100-fold differences were found between gastric carcinoma cell lines and normal tissue cell lines. We also screened bone-marrow RNA of 47 patients with gastric cancers, using this system. Among the three markers we tested, with only about CK20 could we find that 27 of 47 patients were positive.Next, we studied the detection of circulating tumor cells in peripheral blood in patients with gastric cancer using Quantitative RT-PCR. Common mRNA targets for RT-PCR for detection of small numbers of cancer cells in gastric cancer are CK18,CK19,CK20, and CEA. The number of CK19 and CK20 mRNA copies was significantly increased in patients with a non-curative operation or recurrence of gastric cancer (CK19;p=0.0087, CK20;p=0.0022) compared with healthy volunteers. There was a significant difference in tumor stages between CK19 positive and negative patients with curative disease on preoperative diagnosis. Five-year survival rates of patients with CK19 positive or negative cases were 50.0% or 79.0%, respectively (p=0.0347). In conclusion, micrometastases of gastric cancer can be detected in circulating peripheral blood using quantitative real-time RT-PCR. CK19 is a better marker than CK18,CK20 and CEA for peripheral blood, and could be clinically useful to estimate prognosis or to make a postoperative strategy of adjuvant treatment.

实时定量逆转录酶聚合酶链反应（RT-PCR）用于检测骨髓和外周血中的传播癌细胞。首先，我们使用4种胃癌细胞系和3条正常组织细胞系比较了CEA，CK18和CK20表达，以便选择最合适的标记，以检测骨髓中散布的胃癌细胞。事实证明，CK20是最有前途的标记物，因为正常细胞系的表达水平极低，并且在胃癌细胞系和正常组织细胞系之间发现了约50至100倍的差异。我们还使用该系统筛选了47例胃癌患者的骨髓RNA。在我们测试的三个标记中，只有大约CK20，我们可以发现47例患者中有27例是阳性的。我们研究了使用定量RT-PCR检测胃癌患者外周血中循环肿瘤细胞的检测。用于检测胃癌少量癌细胞的RT-PCR的常见mRNA靶标是CK18，CK19，CK20和CEA。与健康的志愿者相比，非耐疗法或复发的患者的CK19和CK20 mRNA拷贝数显着增加（CK19； P = 0.0087，CK20； P = 0.0087，CK20； P = 0.0022）。术前诊断中患有治愈性疾病的CK19阳性患者和阴性患者之间的肿瘤阶段存在显着差异。 CK19阳性或阴性病例患者的五年生存率分别为50.0％或79.0％（p = 0.0347）。总之，使用定量的实时RT-PCR，可以在循环血液中检测到胃癌的微转移。 CK19比CK18，CK20和CEA更好，用于外周血，在临床上可以估计预后或制定辅助治疗的术后策略。