Elevated levels and mitogenic activity of lysophosphatidylinositol in k-ras-transformed epithelial cells

In cell lines stably (KiKi) or reversibly (Ts) transformed by the k-ras oncogene originated from a differentiated rat thyroid line (FRTL5 cells), k-ras-induced transformation has been associated with an increased phospholipase A2 activity. Here we provide evidence that this enzymic activity is phosphoinositide specific and leads to the formation of lysophosphatidylinositol. The levels of this lysolipid increased by 2-3-fold in ras-transformed cells (KiKi cells and Ts cells at the permissive temperature of 33 degrees C) as compared to differentiated cells (FRTL5) or to Ts cells maintained at 39 degrees C, i.e. at the temperature where ras-p21, the product of the ras oncogene, is inactive. Since another lysoderivative, lysophosphatidic acid, has been shown to be a mitogen, we have tested whether lysophosphatidylinositol could have a similar activity on thyroid cells. Lysophosphatidylinositol (10-100 microM) induced a 5-10-fold increase in [3H]thymidine incorporation in both FRTL5 and KiKi cells, whereas lysophosphatidic acid was active only in differentiated cells. Lysophosphatidylinositol (approximately 25 microM) and lysophosphatidic acid (50-100 microM) acted synergistically with insulin in increasing [3H]thymidine incorporation. Moreover, lysophosphatidylinositol at concentrations three-fold higher than those found to be mitogenic, inhibited the activity of the GTPase-activating protein. We conclude that lysophosphatidylinositol is a mitogen that might play a role in the modulation of k-ras transformed cell proliferation.