EFFECT OF MAGNESIUM CONTENT ON DENSITY-DEPENDENT REGULATION OF THE ONSET OF DNA-SYNTHESIS IN TRANSFORMED 3T3-CELLS

  • 1 January 1982
    • journal article
    • research article
    • Vol. 42  (5) , 1761-1768
Abstract
A spontaneously transformed clone of BALB/c 3T3 cells became more transformed after > 90 passages as indicated by increased rounding of cells, multiplication to a higher saturation density and increased ability to form colonies when suspended in agar. When the extracellular concentration of Mg2+ was sharply reduced, the highly transformed cells flattened, assumed the shape of nontransformed cells and became regularly arranged in cohesive arrays. If crowded when deprived of Mg2+ they lost more intracellular Mg2+ than did nontransformed and early passage-transformed cells and remained at constant cell density for at least 10 days. The intracellular content of neither Na+ nor K+ changed consistently with Mg2+ deprivation, but the Ca2+ content increased > 2-fold. The sensitivity of the onset of DNA synthesis to inhibition by Mg2+ deprivation increased with the extent of crowding of the cultures. This was demonstrated by varying population density within a single culture dish as well as from culture to culture. The loss of intracellular Mg2+ in low concentrations of extracellular Mg2+ increased with cell crowding as did the inhibition of DNA synthesis per fractional loss of intracellular Mg2+. Neither deprivation of K+ or Ca2+ nor addition of cAMP produced a density-dependent inhibition of DNA synthesis. A reduction of the Mg2+ content of highly transformed cells apparently restores density-dependent inhibition of the onset of DNA synthesis, which is a characteristic property of nontransformed cells. The differences in Mg2+ retentiveness with population density may reflect differences in intracellular distribution and binding of Mg2+, which could in turn explain some of the regulatory effects of population density on metabolism and growth.