Steck P A, Nicolson G L
Exp Cell Res. 1983 Sep;147(2):255-67. doi: 10.1016/0014-4827(83)90208-2.
Rat 13762NF mammary adenocarcinoma cell surface glycoproteins from s.c. tumor- or lung metastases-derived cell clones of differing spontaneous metastatic potentials were examined for their relationship to metastasis. After treatment with neuraminidase, lectin-binding assays showed that highly metastatic clone MTLn3 cells express approximately twice the quantity of peanut agglutinin (PNA) binding sites (approximately 2.3 X 10(8) sites/cell) than clones of lower metastatic potential. However, the number of wheat germ agglutinin (WGA)-binding sites on the various cell clones decreased slightly as the metastatic potential of the clones increased. The quantities of concanavalin A (conA)-binding sites were similar (approximately 1.7 X 10(8) sites/cell) in all cell clones and growth conditions. Glycoprotein analysis was performed by electrophoresis on polyacrylamide gels in the presence of sodium dodecyl sulfate (SDS-PAGE) and subsequent staining with 125I-labeled lectins. SDS-PAGE gels stained with 125I-labeled conA revealed mainly one glycoprotein (Mr approximately 150 kD), and the amounts of this glycoprotein did not correlate with metastasis. Differences in WGA-binding glycoproteins were detected between s.c. tumor- and lung metastases-derived cell clones. Several desialylated glycoproteins were detected with 125I-labeled PNA after SDS-PAGE, and the labeling intensity of one (Mr approximately 580 kD) correlated with the metastatic potentials of the various cell clones. This high Mr galactoprotein was further analyzed by [3H]glucosamine metabolic labeling, solubilization, sequential gel filtration, and chondroitinase ABC treatment prior to SDS-PAGE. The 580 kD galactoprotein was expressed in increased amounts on the more highly metastatic clones. Chemical labeling of cell surface sialic acid residues using periodate treatment followed by [3H]borohydride reduction showed an additional change in a major sialoglycoprotein (Mr approximately 80 kD), which decreased in labeling intensity on clones of increasing metastatic potential. The results suggest quantitative changes in cell surface glycoproteins rather than major qualitative alterations are associated with differences in the metastatic behavior of 13762NF tumor cell clones.
对源自具有不同自发转移潜能的皮下肿瘤或肺转移瘤的大鼠13762NF乳腺腺癌细胞表面糖蛋白与转移的关系进行了研究。用神经氨酸酶处理后，凝集素结合试验表明，高转移克隆MTLn3细胞表达的花生凝集素（PNA）结合位点数量（约2.3×10⁸个位点/细胞）约为低转移潜能克隆的两倍。然而，随着克隆转移潜能的增加，各种细胞克隆上麦胚凝集素（WGA）结合位点的数量略有减少。在所有细胞克隆和生长条件下，伴刀豆球蛋白A（ConA）结合位点的数量相似（约1.7×10⁸个位点/细胞）。通过在十二烷基硫酸钠存在下进行聚丙烯酰胺凝胶电泳（SDS-PAGE），随后用¹²⁵I标记的凝集素染色进行糖蛋白分析。用¹²⁵I标记的ConA染色的SDS-PAGE凝胶主要显示一种糖蛋白（分子量约150kD），且该糖蛋白的量与转移无关。在源自皮下肿瘤和肺转移瘤的细胞克隆之间检测到WGA结合糖蛋白的差异。SDS-PAGE后用¹²⁵I标记的PNA检测到几种去唾液酸化糖蛋白，其中一种（分子量约580kD）的标记强度与各种细胞克隆的转移潜能相关。在进行SDS-PAGE之前，通过[³H]葡糖胺代谢标记、溶解、连续凝胶过滤和软骨素酶ABC处理对这种高分子量半乳糖蛋白进行了进一步分析。在转移潜能更高的克隆上，580kD半乳糖蛋白的表达量增加。使用高碘酸盐处理随后用[³H]硼氢化物还原对细胞表面唾液酸残基进行化学标记，结果显示一种主要唾液糖蛋白（分子量约80kD）有额外变化，其标记强度在转移潜能增加的克隆上降低。结果表明，细胞表面糖蛋白的定量变化而非主要的定性改变与13762NF肿瘤细胞克隆转移行为的差异有关。
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