Effect of magnesium content on density-dependent regulation of the onset of DNA synthesis in transformed 3T3 cells.
A spontaneously transformed clone of BALB/c 3T3 cells became more transformed after more than 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 more than 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 cyclic adenosine 3':5'-monophosphate produced a density-dependent inhibiton of DNA synthesis. The results indicate that a reduction of the Mg2+ content of highly transformed cells 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 of intracellular distribution and binding of Mg+, which could in turn explain some of the regulatory effects of population density on metabolism and growth.
D000818:Animals; D001345:Autoradiography; D002118:Calcium / Q000378:metabolism; D002452:Cell Count; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000378:metabolism*; D000242:Cyclic AMP / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis*; D004622:Embryo, Mammalian; D008274:Magnesium / Q000378:metabolism* / Q000494:pharmacology; D008275:Magnesium Deficiency; D051379:Mice; D008807:Mice, Inbred BALB C; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism
D004247:DNA; D012964:Sodium; D000242:Cyclic AMP; D008274:Magnesium; D011188:Potassium; D002118:Calcium