纳米免疫磁珠联合RT-nest-PCR法检测胰腺癌外周血微转移的敏感度及特异性  

Examining the micrometastasis in the peripheral blood of the pancreatic cancer

作　　者：胡亮[1] 周家华[1] 余泽前[1] 陶可涛[1] 

机构地区：[1]东南大学附属中大医院肝胆外科,江苏南京210009

出　　处：《东南大学学报（医学版）》2009年第4期287-290,共4页Journal of Southeast University(Medical Science Edition)

基　　金：江苏省科技厅资助项目(BS2002034)

摘　　要：目的:通过胰腺癌外周血微转移模型,探讨纳米免疫磁珠联合RT-nest-PCR检测微转移的敏感度及特异性。方法:将胰腺癌细胞PANC-1稀释分别加入到正常人外周血单核细胞中,再按比例加入Fcλ非特异性受体阻断剂以及上皮细胞EP-CAM抗体,4℃下混匀孵育30 min,经分选柱阳性分选后提取RNA,RT-nest-PCR法检测肿瘤基因人端粒酶亚单位(h-TERT)和癌胚抗原(CEA)。结果:联合纳米免疫磁珠和RT-nest-PCR,平均1×107外周血单核细胞可以检测到1个肿瘤细胞。10例良性疾病对照组均未见表达。结论:免疫磁珠联合RT-nest-PCR是敏感度及特异性都较高的检测微转移的方法,可显著提高肿瘤微转移的早期检测率。Objective To evaluate the sensitivity and specificity of magnetic cell sorting （MACS） examining the micrometastasis in the peripheral blood of the pancreatic cancer. Methods Pancreatic cancer cell line（ PANC-1 ） were cultured in vitro and mixed with PBMC. The malignant cell mixtures were incubated with Fcλ. receptor blocking agent and EP-CAM immune magnetic beads at 4 ℃ for 30 min. After positive MACS screening, RT-nest-PCR was applied to detect CEA- mRNA and h-TERT-mRNA of the enriching cells. Result Using MACS and RT-nest-PCR could detect one circulating cancer cell in 107pBMC. Conclusion Magnetic cell sorting（MACS） combined with RT-nest-PCR is a high sensi- tive and specificity technique for detecting the rate of micrometastasis and can improve the early diagnosis rate of micro- metastasis in peripheral blood in patients with cancer.

关 键 词：胰腺癌 微转移 免疫磁珠 逆转录巢式聚合酶链反应 

分 类 号：R735.9[医药卫生—肿瘤] R446.6[医药卫生—临床医学]