[A study on reversion of the human activated c-Ha-ras-transformed cells].

Two mutant cell lines, R-1 and R-2, have been isolated from EJ/NIH (NIH/3T3 transformed by a human activated c-Ha-ras gene) by treatment with ethyl-methane-sulfonate (EMS). They reveal various characteristics of normal cells, and seem to have reversed to the original cells. Especially, R-1 shows a very flat morphology, complete contact inhibition, anchorage dependent cell growth and no tumorigenecity. Although transformed phenotypes are intensively suppressed, R-1 does not seem to secrete suppressive factors to the parent cell line. The activated c-Ha-ras (EJ-ras) could be detected as a 6.6 kb BamHI fragment in R-1 as well as in EJ/NIH. The level of transcription and translation in R-1 was also the same as that in EJ/NIH. Moreover, the DNA isolated from R-1 cells had a high transforming activity to NIH/3H3, suggesting that R-1 contained the EJ-ras as an activated oncogene. The levels of transcription of c-myc, c-fos, p53 and other ras family genes was unchanged between EJ/NIH and R-1. Further, R-1 cells are resistant to retransformation by EJ-ras, v-src, v-mos and SV40T genes, and DNAs isolated from EJ/NIH or NIH/3T3 could not transform R-1 cells. All these findings suggest that some genes necessary for transformation by EJ-ras, except for c-myc, c-fos, p53 and ras proto-oncogenes, are defective or some inhibitory genes against transformation are enhanced in R-1 cells.