DISCOVERY AND CHARACTERIZATION OF 2 NOVEL HUMAN CANCER-RELATED PROTEINS USING 2-DIMENSIONAL GEL-ELECTROPHORESIS

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 P-actin was found to convert nontumorigenic human fibroblasts to tumorigenicity. Recently, the oncogenic potential of this mutant P-actin was verified independently and shown to alter the metastatic phenotype of human cells in conjunction with the myc and I as 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 P-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.