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在结肠癌转移动物模型中筛选出的转移性小鼠结肠癌细胞系中的细胞表面唾液酸蛋白改变。

Cell surface sialoprotein alterations in metastatic murine colon cancer cell lines selected in an animal model for colon cancer metastasis.

作者信息

Bresalier R S, Rockwell R W, Dahiya R, Duh Q Y, Kim Y S

机构信息

Gastrointestinal Research Laboratory, Veterans Administration Medical Center, San Francisco, California 94121.

出版信息

Cancer Res. 1990 Feb 15;50(4):1299-307.

PMID:2297775
Abstract

Alterations in cell surface proteins and glycoproteins may play a key role in determining the metastatic behavior of tumor cells. The cell surface proteins of a series of related murine colon cancer cells selected in an animal model for colon cancer metastasis (R. S. Bresalier et al., Cancer Res., 47: 1398-1406, 1987) were therefore compared by a variety of biochemical methods. Lactoperoxidase-catalyzed iodination of cell surface proteins followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated quantitative and qualitative differences in the cell surface protein profiles of parental cell line 51B (low metastatic potential) and its metastatic derivatives 51B LiM 5 and 51B LiM 6. Labeling of sialic acid-containing proteins suggested that, in the case of at least four of these proteins (Mr 170,000, 120,000, 95,000, and 55,000), this represented an increase in radioactive labeling of sialoglycoproteins from the metastatic lines. Affinity chromatography of solubilized 125I-labeled cell membrane proteins revealed a 2- to 3-fold increase in wheat germ agglutinin and Sambucus nigra lectin binding associated with the metastatic lines, compared to the poorly metastatic parent. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of material eluted from these columns demonstrated enhancement of proteins from the metastatic cells corresponding in molecular weight to the previously identified major sialoglycoproteins. Neuraminidase-releasable membrane-associated sialic acid and sialyltransferase activities were 2- to 3-fold higher in the metastatic cell lines compared to the parental line. Liver colonization after intrasplenic injection of the various lines into syngeneic mice was dramatically reduced by prior removal of cell surface sialic acid. Immunohistochemical staining of primary and metastatic tumors formed after cecal injection of parental 51B suggested selective metastasis by wheat germ agglutinin-binding tumor cells. These results further support the concept that cell membrane sialylation is important in determining the metastatic potential of cancer cells.

摘要

细胞表面蛋白和糖蛋白的改变可能在决定肿瘤细胞的转移行为中起关键作用。因此,通过多种生化方法比较了在结肠癌转移动物模型中选择的一系列相关小鼠结肠癌细胞的细胞表面蛋白(R.S.布雷萨利尔等人,《癌症研究》,47:1398 - 1406,1987)。用乳过氧化物酶催化细胞表面蛋白碘化,然后进行十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳,结果显示亲代细胞系51B(低转移潜能)及其转移衍生物51B LiM 5和51B LiM 6的细胞表面蛋白谱存在定量和定性差异。对含唾液酸蛋白的标记表明,就这些蛋白中的至少四种(分子量分别为170,000、120,000、95,000和55,000)而言,这代表转移系唾液糖蛋白放射性标记的增加。对溶解的¹²⁵I标记细胞膜蛋白进行亲和层析显示,与低转移亲代相比,转移系与麦胚凝集素和黑接骨木凝集素结合增加了2至3倍。对从这些柱上洗脱的物质进行十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳显示,转移细胞中分子量与先前鉴定的主要唾液糖蛋白相对应的蛋白有所增加。与亲代系相比,转移细胞系中神经氨酸酶可释放的膜相关唾液酸和唾液酸转移酶活性高2至3倍。将各种细胞系经脾内注射到同基因小鼠体内后,预先去除细胞表面唾液酸可显著减少肝内定植。对盲肠注射亲代51B后形成的原发性和转移性肿瘤进行免疫组织化学染色表明,麦胚凝集素结合肿瘤细胞具有选择性转移。这些结果进一步支持了细胞膜唾液酸化在决定癌细胞转移潜能中很重要这一概念。

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