The Effects And Mechanisms Of Glycyrrhetinic Acid On The Invasiveness And Migration Ability Of Leukemic Cells

Objective: To observe the effect of invasiveness and migration ability of leukemia cells treated by Glycyrrhetinic acid(GA),and study the mechanism.Method: The study was performed on cultured K562 and HL-60 leukemia cells, MTT assay was performed to examine the effects of glycyrrhetic acid on the proliferation of cells. The invasiveness of cells were tested by cell matrigel invasion assay. The migration ability of cells were tested by transwell motility assay. The expression of mRNA and protein of MMP-2 and MMP-9 in K562 and HL-60 cells treated by GA were assayed by semi-quantitied RT-PCR and Immunohistochemical studies.Gelatin zymography was used to detect the activity of gelatinase in tested cells.Results : 1. GA showed marked inhibition on proliferation of K562 and HL-60 cells in time-and dose-dependent manner。2.There was statistically significant difference in the inhibition rate of invasiveness and migration ability between the control group and GA groups(30,60,120μmol·L-1).3.GA can downregulate the MMP-2and MMP-9 mRNA level of K562and HL-60 cells. In K562 cells, There was statistically significant difference between the control group and 60μmol·L-1 and 120μmol·L-1 GA groups in expression of MMP-2 mRNA (P<0.05),but 30μmol·L-1 GA groups has no significant difference with control groups (P>0.05). The MMP-9 mRNA level of every groups treated by GA has statistically significant difference with control group(P<0.05).In HL-60 cells, there were statistically significant difference between the control groups and 30,60,120μmol·L-1 GA groups in the mRNA expression of not only MMP-2 but also MMP-9(P<0.05) .4. GA can downregulate the expression of protein of MMP-2 and MMP-9 in treated K562 and HL-60 cells.In K562 cells, There was statistically significant difference between the control groups and 60μmol·L-1 and 120μmol·L-1 GA groups in expression of MMP-2 and MMP-9 protein (P<0.05), but 30μmol·L-1 GA groups has no significant difference with control groups (P>0.05). In HL-60 cells, there were statistically significant difference between the control groups and 60,120μmol·L-1 GA groups in the protein expression of MMP-2 (P<0.05), but 30μmol·L-1 GA groups has no significant difference with control groups (P>0.05). The expression of MMP-9 protein between control and every GA treated groups in HL-60 has statistically significant difference (P<0.05).5. GA can inhibit the activity of gelatinase of the treated cells.There was statistically significant difference between the control group and 60,120μmol·L-1 GA groups.There was only MMP-2 band in K562 cells, but we found brilliant MMP-2 and MMP-9 band in HL-60 cell groups. Both K562 cells and HL-60 cells, there was statistically significant difference between the control groups and 60μmol·L-1 and 120μmol·L-1 GA groups in expression of MMP-2 and MMP-9 (P<0.05),but 30μmol·L-1 GA groups has no significant difference with control groups (P>0.05).Conclusions: GA can inhibit invasiveness and migration ability of the treated K562 and HL-60 cells.Potentially, these effects of GA were achieved through the inhibition of the expression of mRNA and protein, and activity of MMP-2 and MMP-9. This provides the molecular basis for treatment of leukemia patients with this compound.