Experimental Study On Proliferation And Apoptosis Of Human Promyelocytic Leukemia HL-60 Cells Induced By Huaier

Objective To investigate the effects of different concentrations of Huaierqing on proliferation and apoptosis of human promyelocytic leukemia HL-60 cell line..Methods The human promyelocytic leukemia cell line HL-60 was cultured to logarithmic growth phase,in different concentrations of Huaier cream(0mg/ml,2mg/ml,4mg/ml,8mg/ml,16mg/ml),and then cultured in HL-60 cells at 24 h,48h and 72 h.0mg/ml as the control group,2mg/ml,4mg/ml,8mg/ml,16mg/ml as the experimental group.The collecting cell was counted after trypan blue staining to evaluate the effect of Huaierqing cream on the proliferation of HL-60 cells.The absorbance of the cultured cells was measured by CCK-8 cell proliferation assay at 24 h,48h and 72 h to investigate the inhibitory effect of Huaier Qingzhi on proliferation of HL-60 cells.The Annexin V-FITC/PI double staining flow cytometry was used to investigate the effect of Huaier Qingzhi on apoptosis of HL-60 cells.Results 1.After the intervention of Huaier Qing cream,the morphology of HL-60 cells which were observe under the inverted microscope were changed with the increase of drug concentration and the acting time.And the cell density decreased,the refractive index decreased,the cell body shrinked and ruputured.2.The cell counts of HL-60 cells were observed at 24 hours,48 hours and 72 hours after HL-60 cells were treated with different concentrations of Huaier Cream(2,4,8 and 16 mg / ml)respectively,and the results shown that the cell counts in the groups of different concentration of Huaier Qing cream compared with the control group at the corresponding time point decreased,the difference was statistically significant excpt the 2 mg / ml group in which the cells were being acted for 48 hours.3.The results of CCK-8 test showed that the inhibition rates of HL-60 cells were significantly different compared with the negative control group in different concentrations of Huaier cream(2,4,8 and 16 mg / ml)(P <0.001).In a certain concentration range(2-8mg / ml),the inhibition rate was significantly increased with increasing drug concentration and prolonging.There was no statistically significant difference between the inhibition rates of the 16 mg / ml Huaier cream concentration in 48h and 72 h.(P> 0.05).4.Annexin V-FITC / PI double staining flow cytometry: The apoptotic rates of HL-60 cells were higher than the control group at different time(24h,48 h and 72h)in different concentrations(2,4,8 and 16 mg / ml)of the Huaier paste,the difference was statistically significant(P<0.01).Compared with the control group,the apoptotic rate of the experimental group was increased,there has statistically significant difference.And with the prolongation of time and the increase of drug concentration,the apoptosis rate increased gradually,but there was no significant difference between groups(P = 0.0784)?Conclusion 1.Huaier in(2,4,8 and 16 mg / ml)could inhibit the proliferation of HL-60 cells in different concentration range(2-8mg / ml).With the increase of drug concentration,The role of time,the inhibition rate gradually increased.2.Huaier in(2,4,8 and 16 mg / ml)could induce the apoptosis of HL-60 cells at different concentrations,and the low concentration group(2mg / ml and 4mg / ml)Mortality rate increased linearly.In the high concentration group(8mg / ml and 16 mg / ml),the apoptosis rate of 48 hours was significantly higher than that of 24 hours,and the apoptotic rate was lower than that of 48 hours.3.Huaier Qing cream can inhibit the proliferation of HL-60 cells in vitro and promote its apoptosis.