Discovery and characterization of two novel human cancer‐related proteins using two‐dimensional gel electrophoresis
Discovery and characterization of two novel human cancer‐related proteins using two‐dimensional gel electrophoresis
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DOI：
10.1002/elps.1150150154
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发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
J. Leavitt
中科院分区：
文献类型：
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作者：
J. Leavitt
在正常和肿瘤的人成纤维细胞的蛋白质合成的比较检查导致两个新的微丝蛋白质的发现与人类肿瘤的作用。一种蛋白质，一种突变的β-肌动蛋白，被发现可以将非致瘤性人类成纤维细胞转化为致瘤性。最近，这种突变β-actin的致癌潜力被独立验证，并显示与myc和ras癌基因一起改变人类细胞的转移表型。第二种蛋白质，白细胞浆蛋白，被发现是大多数非造血来源的人类癌细胞的标志物。对SV 40转化的人成纤维细胞和人肉瘤和癌细胞类型的调查表明，L-plastin基因在几乎所有人癌细胞中以广泛不同的程度被激活。使用敏感的逆转录聚合酶链反应在正常非造血细胞中未检测到L-plastin基因的激活，除了表达雌激素和孕激素受体的细胞，这些细胞介导生殖组织中L-plastin合成的激活。我们最近的研究结果表明，在不能磷酸化L-plastin的肿瘤细胞（例如仅表达痕量L-plastin的肿瘤细胞类型）中，L-plastin合成的激活导致两种替代性炎症基因表达程序的共诱导，这两种炎症基因表达程序介导对周围细胞的细胞溶解作用。这种炎症反应似乎是由“不适当的”组成型合成的L-plastin和失败的诱导细胞磷酸化L-plastin介导的。我们的研究结果为人类细胞对体外转化的新抗性和癌基因激活在癌症中的作用提供了解释。由于二维（2-D）凝胶电泳分析与其他复杂的分子生物学技术的相互作用，完成了两个基本重要的多基因家族的正式表征，确定了这些蛋白质及其基因的结构和功能的许多方面。突变型β-肌动蛋白和L-plastin的发现和表征及其与人类肿瘤表型的关系可作为使用二维凝胶电泳发现其他重要疾病相关蛋白/基因的有用模型。
Comparative examination of protein synthesis in normal and neoplastic human fibroblasts led to the discovery of two novel microfilament proteins with roles in human neoplasia. One protein, a mutant β‐actin was found to convert nontumorigenic human fibroblasts to tumorigenicity. Recently, the oncogenic potential of this mutant β‐actin was verified independently and shown to alter the metastatic phenotype of human cells in conjunction with the myc and ras oncogenes. A second protein, leukocyte plastin, was discovered to be a marker of a majority of human cancer cells of nonhemopoietic origin. A survey of SV40‐transformed human fibroblasts and human sarcoma and carcinoma cell types demonstrated that the L‐plastin gene was activated at widely varying degrees in nearly all human cancer cells. Activation of the L‐plastin gene was not detected in normal nonhemopoietic cells using sensitive reverse transcript‐polymerase chain reaction, excepting those cells that expressed estrogen and progesterone receptors which mediate activation of L‐plastin synthesis in reproductive tissues. Our most recent findings have revealed that activation of L‐plastin synthesis in neoplastic cells that cannot phosphorylate L‐plastin (e.g. those neoplastic cell types that express only trace amounts of L‐plastin) results in the coinduction of two alternative inflammatory programs of gene expression which mediate cytolytic effects on surrounding cells. This inflammatory response appears to be mediated by “inappropriate” constitutive synthesis of L‐plastin and failure of the induced cell to phosphorylate L‐plastin. Our findings suggest explanations for the novel resistance of human cells to in vitro transformation and one role of oncogene activation in cancer. As a consequence of the interplay of two‐dimensional (2‐D) gel electrophoretic analyses with other sophisticated techniques of molecular biology, the formal characterization of two fundamentally important multigene families was completed with determination of many aspects of the structure and function of these proteins and their genes. The discovery and characterization of the mutant β‐actin and L‐plastin and their relationship to the human neoplastic phenotype serve as useful models for the discovery of other important disease‐related proteins/genes using 2‐D gel electrophoresis.