Magnesium Deficiency Suppresses Cell Cycle Progression Mediated by Increase in Transcriptional Activity of p21Cip1 and p27Kip1 in Renal Epithelial NRK-52E Cells

Lack of magnesium suppresses cell growth, but the molecular mechanism is not examined in detail. We examined the effect of extracellular magnesium deficiency on cell cycle progression and the expression of cell cycle regulators in renal epithelial NRK-52E cells. In synchronized cells caused by serum-starved method, over 80% cells were distributed in G1 phase. Cell proliferation and percentage of the cells in S phase in the presence of MgCl2 were higher than those in the absence of MgCl2, suggesting that magnesium is involved in the cell cycle progression from G1 to S phase. After serum addition, the expression levels of p21(Cip1) and p27(Kip1) in the absence of MgCl2 were higher than those in the presence of MgCl2. The exogenous expression of p21(Cip1) or p27(Kip1) increased the percentage in G1 phase, whereas it decreased that in S phase. The mRNA levels and promoter activities of p21(Cip1) and p27(Kip1) in the absence of MgCl2 were higher than those in the presence of MgCl2. The phosphorylated p53 (p-p53) level was decreased by MgCl2 addition. Pifithrin-alpha, a p53 inhibitor, decreased the p-p53, p21(Cip1) and p27(Kip1) levels, and the percentage in G1 phase in the absence of MgCl2. Rotenone, a mitochondrial respiratory inhibitor, decreased ATP content and increased the p-p53 level in the presence of MgCl2. Together, lack of magnesium may increase p21(Cip1) and p27(Kip1) levels mediated by the decrease in ATP content and the activation of p53, resulting in the suppression of cell cycle progression from G1 to S phase in NRK-52E cells. J. Cell. Biochem. 112: 3563-3572, 2011. (C) 2011 Wiley Periodicals, Inc.