Ca2+ pools and cell growth: arachidonic acid induces recovery of cells growth-arrested by Ca2+ pool depletion

The intracellular Ca2+ pump blocker, thapsigargin, induces emptying of Ca2+ pools and entry of DDT1MF-2 smooth muscle cells into a quiescent G(0)-like growth state. Although thapsigargin blocks pumps essentially irreversibly, high serum (20%) induces appearance of new pump protein, return of functional pools, and reentry of cells into the cell cycle (Waldron, R. T., Short, A. D., Meadows, J.J., Ghosh, T. K., and Gill, D. L. (1994) J. Biol. Chem. 269, 11927-11933). Through analysis of the effects of defined serum components and growth supplements, we reveal here that the factors in serum responsible for inducing recovery of Ca2+ pools and growth in thapsigargin-arrested DDT1MF-2 cells are exactly mimicked by the three essential fatty acids, arachidonic, linoleic, and alpha-linolenic acids. The EC50 values for arachidonic and linoleic acids on growth induction of thapsigargin-arrested cells were the same, approximately 5 microM. Nonessential fatty acids, including myristic, palmitic, stearic, oleic, and arachidic acids, were without any effect. Although not proven to be the active component of serum, levels of arachidonic and linoleic acids in serum were sufficient to explain serum-induced growth recovery. Significantly, arachidonic or linoleic acids induced complete recovery of bradykinin-sensitive Ca2+ pools within 6 h of treatment of thapsigargin-arrested cells. Protein synthesis inhibitors (cycloheximide or puromycin) completely blocked the appearance of serum-induced or arachidonic acid-induced agonist-sensitive pools. The sensitivity and fatty acid specificity of Ca2+ pool recovery in thapsigargin-arrested cells were almost identical to that for growth recovery. No pool or growth recovery was observed with 5,8,11,14-eicosatetraynoic acid, the nonmetabolizable analogue of arachidonic acid, suggesting that conversion to eicosanoids underlies the pool and growth recovery induced by essential fatty acids. The results provide not only further information on the link between Ca2+ pools and cell growth but also evidence for a potentially important signaling pathway involved in inducing transition from a stationary to a proliferative growth state.