The biological effects of antisense mos expression in fibroblasts

In order to determine the physiological significance of c-mos RNA expression in somatic cells, we introduced antisense c-mos under the control of an inducible promoter. NIH/3T3 cells were stably transfected with antisense mos under the control of the mouse mammary tumor virus long terminal repeat (MMTV-LTR). Positive transfectants were selected under G418 conditions. Following selection, NIH/3T3 cells that received the antisense mos failed to form foci, whereas sense mos transfected cells grew normally. Moreover, v-mos-transformed cells were unaffected by antisense mos transfection. Of interest, NIH/3T3 antisense mos transfectants that survived selection were growth-arrested. Nuclear abnormalities and the extrusion of microvesicles containing cellular material were observed in these cells. In order to rescue these cells from growth inhibition, the v-mos gene was introduced into cells by acute infection with Moloney murine sarcoma virus. Following infection, these cells resumed growth and became rapidly transformed. In other experiments, mouse C2 cells stably transfected with antisense mos showed a slower growth rate and gross morphological changes. C2 cells containing antisense mos under the control of mouse metallothionein-1 promoter had a large and flattened morphology and a relatively high percentage (30%) of binucleated cells. Our results indicate that basal level expression of antisense mos (under uninduced conditions) results in either arrested or retarded cell growth. The phenotypes exhibited in both cell lines leads us to suggest that the c-mos expression may play a role in mitotic progression in some somatic cells, in particular affecting cytokinesis.