Experimental Study Effect Of Iron Chelator On Expression Of NF-κBp65 And IκB-a In K562 Cells

ObjectiveTo investigate the influence of survival,growth and the expression of NF-κB and IκB-a inK562 cells by iron chelator deferoxamine(DFO).To explore out a new effective methodof drug treatment for leukemia.Methods(1)The growth state of K562 cells at first to seventh day is determined by Trypanblue.According to the growth state,the growth curve of K562 cells is drawn.(2)Theexperiment is divided into control group(NS);low concentration of DFOgroup(12.5μmol),middle concentration of DFO group(25.0μmol) and high concentrationof DFO group(50.0nmol).(3)The K562 cells are cultured with experimental groups.At0h,12h,24h and 36h,the expression of N F -κB and IκB-a is detected byimmunocytochemistry.The death cell percentage is calculated through the survivalpercentage determined by Trypan blue.Results(1)The growth of K562 cells is slow at 1～3 day,the growth is quick and be in increasedlogarithmic phase at 3～5 day,the growth is in platform state at 5～7 day.(2)The averageoptical of postive area ofN F-κBp65 and IκB-a.①When K562 cells are cultured withdifferent concentrations of DFO for 0h,12h,24h and 36h,the expression of N F-κBp65between 12h/12.5μmol/L group and control group is no difference (P>0.05),but there is difference between others experimental groups and control groups(P<0.05).At the same time point,there is no difference between the 12.5μmol/L/12h group(average optical:180.96±4.88)and control group;there is no difference between the 25.0μmol/L/12h group(average optical:154.78±8.18)and the 50.0μmol/L/12h group(average optical:149.40±5.10),but there is difference between the two groups and the 12.5μmol/L/12h group;there is no difference between the 25.0μmol/L/24h group(average optical:138.35±2.13)and the 50.0μmol/L/24h group(average optical:136.64±1.22),but there is difference between the two groups and the 12.5μmol/L/24h group;at 36h point,between high concentration of DFO group and low concentration of DFO group,the expression of NF-κBp65 is significance decreased in high concentration of DFO group. At the same concentration point,there is no difference between the 24h/25.0μmol/L group(average optical:138.35±2.13)and the 36h/25.0μmol/L group(average optical:136.64±1.22) (P>0.05),but at the other groups,the expression of NF-KBp65 in longer time group is lower than that in shorter time group.(2)The expression of IκB-a between all experimental groups and control groups is difference(P<0.05).At the same time point,there is difference between the lower concentration of DFO group and the higher concentration of DFO group(P<0.05).At the same concentration point,there is no difference between the 12h/12.5μmol/L group(average optical:169.80±3.00)and 24h/12.5μmol/L group(average optical: 166.72±11.57)(P>0.05),but there is difference between the two groups and the 36h/12.5μmol/L group(P<0.05);there is difference between the other groups(P<0.05).(3)When K562 cells are cultured with different concentrations of DFO,the death cell percentages of K562 cells is gradually increasing.At the same concentration point,there is difference between the only 36h/12.5μmol/L group and control group(P<0.05)at 12.5μmol/L point;there are difference between 24h/25.0μmol/L group,36h/25.0μmol/L group and control group(P<0.05) at 25.0μmol/L point;there are difference between every experimental group and control group at 50.0μmol/L point.At the same time point,there is difference between the only 50.0μmol/L/12h group and control group(P<0.05)at 12h point;there are difference between25.0μmol/L/24h group,50.0μmol/L/24h group and control group(P<0.05) at 24h point;there are difference between every experimental group and control group at 36h point. Conclusions(1)The iron chelator(DFO) can depress the growh of K562 cells. (2)The iron chelator(DFO) can downregulate the expression of N F-κB and IκB-a of K562 cells.(3)It is hopeful to be a safe and effective inhibitor of NF-κB activity and become an alternation in treatment of leukemia and other cancer,but the safety evaluation of DFO in clinical application is required to research deeply.