Inufusa H, Kojima N, Yasutomi M, Hakomori S
Biomembrane Institute, Seattle, WA 98119.
Clin Exp Metastasis. 1991 May-Jun;9(3):245-57. doi: 10.1007/BF01753728.
Human lung adenocarcinoma sub-cell lines HAL-8, HAL-24 and HAL-33, showing different lung colonization potential (LCP), were established from human lung adenocarcinoma cell line KUM-LK-2 using repeated cloning with limiting dilution technique. Cell lines HAL-8 and -33 were characterized by high and low LCP, respectively, while HAL-24 did not give rise to lung colonies. The cell surface protein and carbohydrate profiles were determined by cell surface labeling (with lactoperoxidase-dependent 125I-iodination and galactose oxidase-NaB3H4, respectively) followed by SDS-gel electrophoresis. Various carbohydrate epitopes expressed at the cell surface were analysed by cytofluorometry using various monoclonal antibodies (MAbs) directed to Le(x), sialosyl-Le(x), sialosyl dimeric Le(x), T, Tn and sialosyl-Tn structures, which are often reported as being highly expressed in a variety of human cancers, particularly adenocarcinoma. Expression of sialosyl dimeric Le(x) (defined by MAb FH6) was high on HAL-8, moderate on HAL-33, and relatively low on HAL-24. In contrast, each of the three lines showed essentially equal expression (as determined by MAb reactivity) of sialosyl-Tn (defined by MAb TKH2), Le(x) (defined by MAb SH1), and Tn (defined by MAb 1E3). The cell lines showed extremely weak expression of T (defined by MAb HH8). LCP of HAL-8 and -33 was completely inhibited by sialidase treatment of cells. It is suggested that higher expression of sialosyl dimeric Le(x) (defined by MAb FH6) in HAL-8 cells may play an important role in higher potential of blood-borne lung colonization.
利用有限稀释技术进行重复克隆,从人肺腺癌细胞系KUM-LK-2建立了具有不同肺定植潜能(LCP)的人肺腺癌亚细胞系HAL-8、HAL-24和HAL-33。细胞系HAL-8和HAL-33分别具有高LCP和低LCP特征,而HAL-24未形成肺集落。通过细胞表面标记(分别采用依赖乳过氧化物酶的125I碘化和半乳糖氧化酶-NaB3H4),随后进行SDS凝胶电泳,测定细胞表面蛋白和碳水化合物谱。使用针对Le(x)、唾液酸化Le(x)、唾液酸化二聚体Le(x)、T、Tn和唾液酸化Tn结构的各种单克隆抗体(MAb),通过细胞荧光测定法分析细胞表面表达的各种碳水化合物表位,这些表位在多种人类癌症尤其是腺癌中常被报道为高表达。唾液酸化二聚体Le(x)(由MAb FH6定义)在HAL-8上高表达,在HAL-33上中等表达,在HAL-24上相对低表达。相反,这三个细胞系对唾液酸化Tn(由MAb TKH2定义)、Le(x)(由MAb SH1定义)和Tn(由MAb 1E3定义)的表达(由MAb反应性确定)基本相等。这些细胞系对T(由MAb HH8定义)的表达极弱。用唾液酸酶处理细胞后,HAL-8和HAL-33的LCP被完全抑制。提示HAL-8细胞中唾液酸化二聚体Le(x)(由MAb FH6定义)的高表达可能在血源性肺定植的较高潜能中起重要作用。