The Effect Of Amiloride On Myeloid Leukemia Cells

Objective: This study was aimed to explore the effect of amiloride on the proliferation of acute myeloid leukemia cell line HL-60 cells and chronic myeloid leukemia cell line K562 cells and K562 / ADM cells,and its mechanism.Methods: Cell counting assay detected if the cells were in the logarithmic growth phase; The expression of P-gp was detected by the flow cytometry in the K562 cells and the K562 / ADM cells;The influence of cell proliferation was detected of the amiloride on HL-60 cells, K562 cells and K562 / ADM cells by MTT assay; The m RNA expression of u PA, u PAR, MMP-9 and the c-myc was detected of the amiloride on K562 cells by real-time PCR;The protein expression of u PA, u PAR, MMP-9 and c-myc was detected of the amiloride on K562 cells by Western Blotting.Results: 1 MTT assay showed that amiloride(1×10-3mol/L、1×10-4mol/L、1×10-5mol/L、1×10-6mol/L) could inhibit the proliferation of HL-60 cells, K562 cells and K562 / ADM cells,which was significantly,and it appeared a correlation of time-dependent and dose-dependent, which was significant different compared with the control group.The inhibition increased with the increasing time and doses control group, the difference was significant compared with the control group(P <0.05).2The inhibition was weaker of amiloride on the proliferation of K562 / ADM cells than the proliferation of HL-60 cells and K562 cells(P <0.01).3 The real-time PCR showed that the m RNA expression of u PA, u PAR, MMP-9 and the c-myc decreased significantly when we when we cultured K562 cells with amiloride(1×10-3mol/L)for 24 hours(P <0.05).4Western Blotting showed that the protein expression of u PA, u PAR, MMP-9 and c-myc was significantly lower compared with the control group when we cultured K562 cells with amiloride(1×10-3mol/L) for 24 hours(P <0.05).Conclusion: Amiloride could inhibit the proliferation of leukemia cells, which may be associated with the down-regulation of the expression of u PA,u PAR, MMP-9 and c-myc.