高及低转移潜能小鼠肝癌细胞株糖蛋白凝集素结合特征的研究

为探讨肿瘤转移与细胞表面的糖结构的关系，对小鼠肝癌细胞的高、低淋巴道转移株Ｈｃａ－Ｆ和Ｈｃａ－Ｐ进行了蛋白质电泳及经蛋白质印迹术后的５种凝集素（ＣｏｎＡ、ＷＧＡ、ＵＥＡ、ＳＢＡ、ＰＮＡ）结合糖蛋白谱的对比分析．结果表明：高、低转移两株细胞的ＳＤＳ－ＰＡＧＥ谱基本相同；ＣｏｎＡ特异结合糖蛋白共有５种（～７２，８０～９０，～１０４，～１５０，～２００ｋＤ）；其中较明显的差异为～７２ｋＤＣｏｎＡ特异结合糖蛋白，它在Ｈｃａ－Ｐ细胞的表达明显高于Ｈｃａ－Ｆ细胞．ＷＧＡ特异结合糖蛋白１种（～１５０ｋＤ），在Ｈｃａ－Ｐ细胞的表达略高于Ｈｃａ－Ｆ细胞．此外，实验发现两种性质未明的蛋白质（～７９，～１３０ｋＤ），后者在Ｈｃａ－Ｐ细胞的含量明显高于Ｈｃａ－Ｐ细胞．结果提示Ｈｃａ－Ｆ和Ｈｃａ－Ｐ细胞不同的转移表型可能与其糖蛋白的表达有一定的关联．

Lectin-binding Characteristics of Glycoproteins of Related High-and Low-Metastatic Murine HCa Cell Lines

Two of recently isolated murine hepatocarcinoma subclonal cell ines,HCa-F and HCaP with high-and low-metastatic potential in lymphatic system respectively, were adapted in this study. Cellular glycoproteins of the two cells were analyzed for their relationship to metastasis.After separated by SDS-PAGE and electroblotted to NC membranes,the total cellular glycoproteins were overlaid bv five different biotinlabelled lectins (WGA, Con A, PNA, UEA, SBA) and then revealed by the avidin-biotin peroxidase complex (ABC) method. No apparant differnce of protein separated by SDS-PAGE could be seen between the two cells. The major lectin-binding glycoproteins of the cells detected are: five of Con A-binding glycoproteins of Mr ￣72 . 80 ￣90,￣104, ￣150,￣200 kD which could be blocked by mannose: one of ￣150 kD WGA binding glycoprotein blocked by GLcNAc.There were no specific SBA-UEA-or PAN-binding glycoproteins in these cells. Most of above detected glycoproteins were similar in amout between the highand low-cell lines except that the ￣ 72 kD Con A binding-band, which expressed mainly on HCa-P. This difference might be the result of oligosacchanide changes of glycoprotein. Unexpectedly, two “avidin-binding”proteins of Mr￣79 kD and ￣1 30 kD were detectable in total cellular extracts,and the amount of the latter in HCa-F cells was apparantly higher than that in HCa-P cells. The results suggested that quantitative rather than qualitative changes in cell surface are associated with the difference in metastatic behavior of murine HCa cells.