凝集素芯片对体液细胞进行检测方法的建立和初步应用

Establishment of a Lectin Microarray for the Analysis of Body Fluid Cells and Its Application

建立对体液细胞进行自动捕获的凝集素芯片体系,利用凝集素对糖链的特异亲和作用捕获细胞,提取白血病患者外周血、肺癌胸水和肝腹水中细胞进行荧光标记,凝集素芯片捕获,激光扫描仪检测捕获细胞的荧光信号,常规HE染色后光学显微镜下观察细胞的形态并进行免疫化学反应,流式细胞仪验证凝集素芯片的特异性.结果表明:凝集素芯片可以对体液中的癌细胞进行自动捕获,对癌细胞膜表面糖链进行识别,芯片检测的细胞浓度最少可达每mL 104个左右,芯片有较好的重复性和特异性.这种凝集素芯片可用于临床体液中癌细胞的检测分析,对癌细胞膜表面凝集素亲和位点进行即时、高通量的检测,为了解细胞膜表面聚糖在癌变过程中的变化提供了一个技术平台.

The technology of lectin microarrays was established for catching cells automatically, cells were extracted from peripheral blood, pleural fluid and ascitic fluid of cancer patients and labeled by AO （acridine orange）, and cells were bounded on the slide by the affinity interaction between the lectins on the slide and the glycans on the cell surface, bound cells were directly detected by a laser fluorescence scanner and observed under microscope after stain with HE or immunochemistry reactions. Flow cytometry was used to analyze the specificity of lectin microarray. The consequence of experiment indicated that the lectin microarray can automatically catch malignant cells extracted from body fluid, and the profile of glycome of the cell surface can be obtained. A relatively low number of cells （the liminal concentration is 1 ×10^4）is require and the lectin microarray have good specificity. Hence the lectin microarray can be used to clinical analysis of cancer cells and provides a novel strategy for studying profiling changes of the cell surface glycome on tumor metastasis.