OVER-EXPRESSION OF THE S13 RIBOSOMAL-PROTEIN IN ACTIVELY GROWING CELLS

In order to define biological markers of aggressiveness, 2 rat colon-carcinoma cell lines differing by their tumorigenicity were used to clone genes over-expressed in colon carcinoma as compared with normal epithelial cells. A progressive rat colon-carcinoma clone (PROb) cDNA library was hybridized with 32P-cDNA synthesized from mRNA prepared from these PROb cells, or from regressive cells (REGb) derived from the same tumor. Several clones were isolated after the initial screening. The specificity of each clone was confirmed by RNA blotting. One of these (B9) was found to hybridize to an mRNA 30-fold more abundant in PROb cells than in normal adult rat colon, and was therefore selected for further study. No gene amplification was detected by Southern blot analysis, indicating that the difference in mRNA content was most likely due to an increased transcription of this gene. Sequencing of the cDNA revealed approximately 98% homology with the rat S13 ribosomal protein. The expression level of this gene was determined in a series of rat cell lines with different growth rates. A good correlation was found between these 2 parameters. Our data suggest that the S13 ribosomal-protein gene can be used to evaluate the growth rate of tumor cells, which might be correlated with their aggressiveness. In an initial trial experiment, S13 ribosomal-protein mRNA was detected in a series of human colorectal tumors by in situ hybridization. A strong signal was seen in the 4 tumors analyzed. (C) 1993 Wiley-Liss, Inc.