Effect of potassium bisperoxo (1,10-phenanthroline) oxovanadate on proliferation and cell cycle in HT22 cells / 中国药理学与毒理学杂志

ABSTRACT

OBJECTIVE To explore whether different concentrations of potassium bisperoxo (1,10-phenanthroline) oxovanadate [BPV (phen)) affect cell cycle by regulating the expression of DNA meth-yltranferases (DNMT) and cell cycle-related genes. METHODS HT22 cells were treated with BPV (phen) 0.3 and 3 μ mol • L-1 for 24 h. The cell viability was detected by MTS assay, cell cycle was detected by flow cytometry, the activity of DNMT was detected by ELISA, the mRNA expressions of p21, DNMT1, DNMT3A and DNMT3B were measured with real-time quantitative PCR, while the levels of corresponding proteins were measured by Western blotting. RESULTS Compared with the DMSO control group, BPV(phen) 0.3 μ mol• L-1 did not affect cell viability, but the cell viability of BPV(phen) 3 pmol-L-1 increased signifi-cantly (P<0.05). There was no significant difference in cell cycle between DMSO control and BPV(phen) 0.3 nmol-L-1 group, but the proportion of cells in S phase increased((76.0±1.6)%)(P<0.05) and in G2 phase decreased ((2.1 ±1.5)%) (P<0.05) of BPV(phen) 3.0 μ mol • L-1 group. The intracellular DNMT activity of BPV(phen) 3.0 Mmol • L-1 group was significantly increased compared with the DMSO control group (F<0.05). There was no significant difference in the mRNA expressions of p21, DNMT1, DNMT3A and DNMT3B between DMSO control and BPV(phen) 0.3 μ mol • L-1 group, but all increased in BPV(phen) 3.0 μ mol • L-1 group (P<0.05, P<0.01). Western blotting results showed that there was no significant difference in the protein expressions of P21, DNMT1, DNMT3A and DNMT3B between DMSO control and BPV (phen) 0.3 μ mol • L-1 group, but only the protein expressions of DNMT3B and P21 of BPV (phen) 3.0 μ mol • L-1 group increased significantly (F<0.05). CONCLUSION BPV(phen) can regulate the expression of downstream and cell cycle-related genes by altering the expression of DNMTs, which in turn affects the growth and proliferation of HT22 cells.