磁激活细胞分选术联合混合抗体提高模拟恶性腹水中游离癌细胞检出效率的分析  

Magnetic activated cell sorting combined with a panel of monoclonal antibodies in detecting free cancer cells in analogue malignant ascites

作　　者：王晓蕾[1] 陈锡美[1] 黄志刚[1] 王韶英 

机构地区：[1]上海市同济大学附属同济医院消化科,200065

出　　处：《中华消化杂志》2008年第6期392-396,共5页Chinese Journal of Digestion

基　　金：上海市自然科学基金资助项目（05ZR14130）

摘　　要：背景磁激活细胞分选术（magnetic activated cell sorting，MACS）是一种新的免疫磁性分离技术。其原理是基于抗体对抗原的特异性识别，将50nm磁性微珠直接或者间接耦联在抗体上，与有相应抗原表达的细胞相连，在高强度、高梯度磁场中达到细胞磁性分离的目的。该法具有分离纯度和回收率均较高的优势，也能分离出体液中存在的少量肿瘤细胞。目的评价MACS联合一组肿瘤细胞标志物的方法对提高模拟恶性腹水中游离癌细胞检出效率的作用。方法选择5种与恶性腹水病因有关的肿瘤细胞株作为研究对象，采用免疫荧光反应和流式细胞仪（FCM）检测上皮相关抗原（epithlial-related antigen，EpCAM）、CA125、癌胚抗原和TAG-72共4种单克隆抗体及其混合抗体在各肿瘤细胞的表达。将肿瘤细胞以不同比例掺入单个核细胞中模拟恶性腹水细胞成分，与联合单个抗体进行分选对比，观察MACS术联合混合抗体分选肿瘤细胞的效率。结果FCM结果表明，混合抗体在5种肿瘤细胞的阳性表达率均高于4种抗体单独反应的阳性率。MACS术联合混合抗体检出模拟恶性腹水中肿瘤细胞的得率比联合单个抗体的得率要高，其中以2种胃癌细胞和结肠癌细胞的平均得率提高较多（分别为69．18％±20．84％比45．23％±11．54％、78．75％±15．42％比59．73％±16．640和85．63％±12．30％比76．88％±8．650），卵巢癌细胞次之（32．490±3．58％比31．79％±4．820），肝癌细胞最少（11．78％±0．430比7．16％±0．460）。结论与联合单个抗体进行分选相比，应用MACS术联合一组混合抗体的方法可有效提高恶性腹水中游离癌细胞的检出效率，尤其对胃肠道肿瘤所致的恶性腹水有潜在的临床应用价值。Background Magnetic activated cell sorting （MACS） is a new immunomagnetic separation technique. It＇s principle is based on the specialized recognition between the antibody and antigen. When directed or indirected coupled with the 50 nm magnetic beads, the antibody can linked with the cells which express the corresponding antigen, and then leads to the magnetic cell separation in a high- intensity and high-gradient magnetic field. The method has higher separative purity and recovery and been made use of enriching tumor cells and tumor stem cells. It could be also utilized to enrich the rare tumor cells in fluid specimen. Objective To evaluate the enriching efficiency of detecting free cancer cells in analogue malignant ascites using MACS and a panel of tumor- specific markers. Melhods Five species of tumor cell lines correlated to the diseases resulting in malignant ascites were selected and the expressions of EpCAM, CA125, CEA, TAG-72 and their mixture in these tumor cell lines were detected using immunofluorescence reactions and flow cytometry（FCM）. The tumor cells were spiked into mononuclear cells by different ratios to analogue the main ingredients of malignant ascites. The efficiency of MACS combinded with mixture antibodies in separating tumor cells was compared with single antibody.Results The FCM revealed that the expression of the mixture antibodies in 5 tumor cancer cell lines was significantly higher than that in single antibody. The mean recovery rates of spiked tumor cells were enhanced by using a panel of monoclonal antibodies combined with MACS than single antibody, especially in two gastric cancer cells and colon cancer cells （69. 18% ± 20.84 % vs 45. 23% ±11.54%, 78. 75% ±15.42% vs 59.73±16.61% and 85. 63%±12. 30% vs 76.88% ± 8.65%,respectively）, the ovarian cancer cells was the next （32.19%±3. 58% vs31. 79%±4.82%）, and the liver cancer cells was the lowest （11.78%±0.43% vs 7. 16%±0.46%）. Conclusions The detection of free cancer cells from malignant ascites by MACS

关 键 词：肿瘤转移 腹水液 免疫磁化分离 磁激活细胞分选术 

分 类 号：R686[医药卫生—骨科学]