摘要
目的建立定量检测小细胞肺癌（SCLC）循环肿瘤细胞（CTC）的方法，分析其敏感性和稳定性。方法设计前胃泌素释放肽原（preproGRP）特异性引物和探针，建立检测preproGRPmRNA的定量逆转录聚合酶链反应（RT-PCR）方法，通过癌细胞掺入法评价检测的敏感性。同时采用免疫磁珠法（MACS）分离纯化外周血CTC，结合形态学诊断计数CTC。结果MACS分离外周血CTC的敏感性为30％左右，检出下限为5个CTC／ml全血。preproGRP定量RT-PCR检测外周血CTC的敏感性为0．64个CTC／反应，检出下限为50个CTC／ml全血，低于MACS，但通过Ct值计算的CTC数量与实际细胞数量符合程度（80％左右）高于MACS法。结论preproGRP定量RT-PCR和MACS在检测SCLC患者外周血CTC方面各有优缺点，MACS敏感性更高，适合〈50个CTC／ml全血的检测，preproGRP定量RT-PCR检测的CTC数量更符合实际含量。
Objective To establish a quantitative method to detect circulating tumor cells （CTC） in patients with small cell lung cancer, and analyze its sensitivity and stability. Methods A specific primer and probe for prepro-gastrin-releasing peptide （preproGRP） was designed and a quantitative RT-PCR method was established to detect preproGRP mRNA. Cell incorporation method was used to evaluate the sensitivity. Magnetic cell sorting （MACS） was used to isolate and purify CTC from peripheral blood, and the MACS in combination with morphological diagnosis were used for cell counting. Results The isolation rate of CTC by MACS was 30% and the lower detection limit was 5 cells per ml blood. The sensitivity of quantitative RT-PCR in detection of preproGRP mRNA in CTC was 0.64 ceils per reaction, and the lower detection limit was 50 cells per ml blood, which was lower than that of MACS. However, the cell numbers calculated by Ct value was in greater accordance （about 80% ） with actual cell numbers than that obtained by MACS. Conclusions PreproGRP quantitative RT-PCR and MACS have both advantages and disadvantages in detecting CTC of SCLC patients. MACS has a higher sensitivity, and is more favorable when CTC count is below 50 per ml blood. Meanwhile, preproGRP mRNA quantitative RT-PCR is more reliable in calculating actual cell numbers.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2013年第5期347-350,共4页
Chinese Journal of Oncology
基金
山东省医药卫生科技发展计划（2005HZ089）
关键词
癌
小细胞
前胃泌素释放肽原
免疫磁珠法
循环肿瘤细胞
血液
Carcinoma, small cell
Prepro-gastrin-releasing peptide
Magnetic cell sorting
Circulating tumor cells, blood
