H-RAS EXPRESSION, GENETIC INSTABILITY, AND ACQUISITION OF METASTATIC ABILITY BY RAT PROSTATIC-CANCER CELLS FOLLOWING V-H-RAS ONCOGENE TRANSFECTION

To study the relationship between metastatic ability, mutated H-ras expression, and genetic instability, a cloned, nonmetastatic rat prostatic cancer cell line (AT2.1) was transfected with the v-H-ras oncogene. The parental AT2.1 clone, 4 control transfectants (Neo/Only), and 9 v-H-ras transfectants (Neo/Ras) were characterized with regard to their H-ras content by using Southern, Northern, and Western blot analysis and their biological behaviour in vivo. Following s.c. inoculation in syngeneic rats, all transfectants produced tumors. All 4 (Neo/Only) transfectants like the parental untransfected cell were non-metastatic. Six of 9 Neo/Ras transfectants were metastatic to the lungs and lymph nodes, while the other 3 Neo/Ras transfectants were not metastatic. There was no simple dose-response relationship between the level of v-H-ras integration, mRNA or p21 protein expression, and the development of metastatic ability by the Neo/Ras transfectants. Cytogenetic analysis demonstrated that the frequency of additional structural and/or additional numerical chromosomal changes among the Neo/Ras transfectants was significantly higher than that in the Neo/Only transfectants (P < 0.05). Loss of chromosome 10 was observed in all of the Neo/Ras transfectants, whereas that was observed in only one of the 4 Neo/Only transfectants (P < 0.05). There were no specific chromosomal changes, however, which were statistically correlated with the development of metastases in the Neo/Ras transfectants. These results demonstrate that development of the metastatic ability in AT2.1 cells is not a single-step reaction regulated by the level of H-ras expression alone, but rather a complex process requiring additional events. One of the additional events appears to be an increase in genetic instability and cytogenetic changes following v-H-ras transfection.