Effects Of Mangiferin On Expression Of FLT3 Wild-type And Mutant Acute Myeloid Leukemia Cells

Objective: To investigate the effects of mangiferin on proliferation,cell cycle,apoptosis,FLT3 mRNA expression and FLT3 transmembrane protein expression in FLT3-ITD mutant AML cell line MV4-11 and FLT3 wild type AML cell line THP-1.To provide a new theoretical basis for the development of mangiferin as a new drug for the treatment of leukemia.Methods: AML cell line MV4-11 expressing FLT3-ITD mutation positive and AML cell line expressing THP-1 expressing FLT3 wild type were selected as study subjects,and PBS blank control was used to treat mangiferin with different concentrations(0,0.25,0.50,0.75,1.00,1.25 mmol/L)MV4-11 and THP-1 cells were treated separately for different times(12 hours,24 hours,48 hours,72 hours,96 hours).The CCK-8 method was used to determine the proliferative activity of mangiferin at different concentrations for different time;the half-inhibitory concentration(IC50)of cell proliferation was calculated according to the results of CCK8 experiment,and the optimal conditions for mangiferin were screened,and the concentration and time were selected.The concentration and time of the cells were treated as subsequent experiments.The effects of different concentrations of mangiferin on the apoptosis and cell cycle of FLT3-ITD mutation-positive AML cell line MV4-11 and FLT3 wild-type THP-1 cells were detected by flow cytometry.Effects of mangiferin on the expression of FLT3 transmembrane protein in MV4-11 and THP-1 cells for 48 h;real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the effects of different concentrations of mangiferin on MV4-11 and THP-1.The expression level of FLT3 mRNA after 48 h of cells.Results:1.CCK-8 assayed the effect of different concentrations of mangiferin on the proliferation inhibition of FLT3-ITD mutant positive AML cell line MV4-11 and FLT3 wild type AML cell line THP-1.The results showed that mangiferin significantly inhibited MV4-11 and THP.-1 cell proliferation,and with the increase in the concentration and duration of action of mangiferin,the inhibition of proliferation of MV4-11 and THP-1 cells was more pronounced,concentrationand time-dependent.2.Different concentrations of mangiferin induced early apoptosis of FLT3-ITD mutation-positive AML cell line MV4-11 and FLT3 wild-type AML cell line THP-1,and the apoptosis was more obvious with the increase of mangiferin concentration.3.Flow cytometry was used to detect the effects of different concentrations of mangiferin on the cell cycle of FLT3-ITD mutation-positive AML cell line MV4-11 and FLT3 wild-type AML cell line THP-1.The results suggest that mangiferin can block MV4-11 and THP.-1 cell cycle in G0/G1 to S phase.4.Real-time quantitative PCR was used to examine the effects of different concentrations of mangiferin on the expression of FLT3 mRNA in FLT3-ITD mutant AML cell line MV4-11 cells and FLT3 wild type AML cell line THP-1.The results showed that mangiferin significantly inhibited the expression of FLT3 mRNA in MV4-11 cells and THP-1 cells,and inhibited the expression of THP-1 in FLT3 wild-type AML cells.MV4-11 cells and THP-1 cells were treated with PBS and PBS.There were significant differences in the control group(P<0.05).5.Flow cytometry detection of IC50 concentration of mangiferin FLT3-ITD mutation-positive AML cell line MV4-11 and FLT3 wild-type AML cell line THP-1 for 48 h,MV4-11 cells and THP-1 cell FLT3 transmembrane The effect of protein expression showed that the expression rate of FLT3 transmembrane protein in mannose-glycoside group of MV4-11 cells and THP-1 cells was significantly lower than that of PBS control group,and the difference was statistically significant compared with PBS control group.(P<0.05).Conclusion:1.Mangiferin inhibited the proliferation of FLT3-ITD mutant cell line MV4-11 and FLT3 wild-type cell line THP-1,and it was concentration-time dependent in a certain concentration range.2.Mangiferin promoted early apoptosis of FLT3-ITD mutant cell line MV4-11 and FLT3 wild-type cell line THP-1,leading to cell cycle G0/G1 arrest.3.Mangiferin can down-regulate the expression of FLT3 gene in MV4-11 and THP-1 cell lines and inhibit FLT3 activity,which may be one of the reasons why mangiferin interferes with AML cell proliferation,induces apoptosis and blocks the cycle.