肿瘤与正常细胞表面糖链结构的流式细胞术分析

Flow Cytometric Analysis of the Sugar Chains on the Surface of Normal and Human Hepatocellular Carcinoma Cells

为了比较正常与肿瘤细胞表面的糖链结构差异,凝集素用荧光素标记后作为研究细胞膜糖链结构的探针,采用流式细胞技术在分子水平上分析。结果显示正常与肿瘤肝细胞与同一浓度的凝集素探针结合量有明显的不同。与凝集素ConA-FITC结合后,正常肝细胞的荧光峰较肿瘤肝细胞的荧光峰明显右移;与凝集素WGA-FITC、PHA-FITC结合后,正常肝细胞的荧光峰较肿瘤肝细胞的荧光峰明显左移。由于凝集素可识别特定糖链结构,该方法说明肿瘤肝细胞的糖链结构与正常肝细胞相比具有特征变化,即平分型糖链和唾液酸含量丰富,可能出现偏二天线以及天线数的增加。

Lectins were fluorescein-labeled to detect the structure of oligosaccharides. By use of the flow cytometry technology, the structure of oligosaccharides from the surface of normal and human hepatocellular carcinoma cells were compared. The mean intensity of fluorescence (MIF) of the normal and human hepatocellular carcinoma cells were determined and calculated after simultaneously incubated with ConA-FITC, WGA-FITC, PHA-FITC and LCA-FITC at the same concentration respectively. There were obvious differences between the two cell lines. The MIF of the normal and carcinoma cells were obviously different. After incubated with ConA-FITC, the MIF of the normal hepatocytes was stronger than that of the carcinoma cells. But compared with human hepatocellular carcinoma cells, the fluorescence peak of normal hepatocytes shifted left after incubated with WGA-FITC, PHA-FITC and LCA-FITC, the MIF of the normal hepatocytes was less than that of the carcinoma cells. There were extraordinary changes found in carcinoma cells compared with that of normal hepatocytes, with the increase of the amount of antennary, complex-type oligosaccharides, aberrant bi-antennary N-glycan and notably increased sialylation.