Marks A, Petsche D, O'Hanlon D, Kwong P C, Stead R, Dunn R, Baumal R, Liao S K
Banting and Best Department of Medical Research, University of Toronto, Canada.
Exp Cell Res. 1990 Mar;187(1):59-64. doi: 10.1016/0014-4827(90)90116-r.
The synthesis of S100 protein in cultured human melanoma cells was examined using metabolic labeling with [35S]methionine, immunoprecipitation with anti-S100 protein antiserum, and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Six of seven cell lines derived from melanomas synthesized relatively large amounts of S100 protein, whereas three cell lines derived from normal melanocytes synthesized lesser amounts. Synthesis of S100 protein was not detected in 10 human cell lines of nonneuroectodermal origin. Analysis of poly(A+) RNA from one melanoma cell line by Northern blot hybridization with a probe specific for the beta subunit of rat S100 protein revealed a single mRNA species of 1.0 kb coding for the human protein. Flow cytometric analysis of individual cells of two melanoma cell lines and the rat glioma cell line C6 indicated that G0/G1 cells were heterogeneous with respect to S100 protein expression, while almost all the cells in S + G2 + M expressed S100 protein. These results suggest that expression of S100 protein in G0/G1 could be a prerequisite for progression of the cells through the cell cycle.
利用[35S]甲硫氨酸进行代谢标记、用抗S100蛋白抗血清进行免疫沉淀以及在十二烷基硫酸钠存在下进行聚丙烯酰胺凝胶电泳,对培养的人黑色素瘤细胞中S100蛋白的合成进行了检测。源自黑色素瘤的七个细胞系中有六个合成了相对大量的S100蛋白,而源自正常黑素细胞的三个细胞系合成量较少。在10个非神经外胚层来源的人细胞系中未检测到S100蛋白的合成。用大鼠S100蛋白β亚基特异性探针通过Northern印迹杂交分析一个黑色素瘤细胞系的聚腺苷酸加尾(poly(A+))RNA,发现有一个1.0 kb的单一mRNA种类编码人蛋白。对两个黑色素瘤细胞系和大鼠胶质瘤细胞系C6的单个细胞进行流式细胞术分析表明,G0/G1期细胞在S100蛋白表达方面存在异质性,而S + G2 + M期的几乎所有细胞都表达S100蛋白。这些结果表明,G0/G1期S100蛋白的表达可能是细胞通过细胞周期进展的一个先决条件。