Effect And Possible Mechanism Of The Flavonoids From Hedyotis Diffusa Willd And Cisplatin On Proliferation And Apoptosis Of Lung Cancer A549 Cells

Objectives:Lung cancer A549 cells were used as research objects in order to investigate the effects of the flavonoids from Hedyotis diffusa willd and cisplatin on proliferation and apoptosis of lung cancer A549 cells and possible mechanisms.Methods:Lung cancer A549 cells were cultured in vitro and logarithmic phase tumor cells were used for this experiment.They were randomly divided into four experimental groups: negative control group;the flavonoids from Hedyotis diffusa willd;cisplatin group;combination group(the flavonoids from Hedyotis diffusa willd and cisplatin group).MTT assay was used to detect the effects of the combined use of the flavonoids from Hedyotis diffusa willd and cisplatin alone and their combination on the proliferation of lung cancer A549 cells,and the Q value formula was used to evaluate the effect of the combined intervention of the two drugs;Cellular analyzers were used to examine and determine the effect of two intervention drugs on the differentiation cycle of cells and the apoptotic rate of cells;RT-PCR was used to detect the expression of apoptosis-related proteins p-AKT,Caspase-9,Bcl-2,and Bax m RNA.Western blotting was used to detect the expression of p-AKT,Caspase-9,Bcl-2 and Bax protein.Results:1.Compared with the negative control group,the flavonoids from Hedyotis diffusa willd and cisplatin groups exhibited time and concentration-dependent inhibition of the proliferation of lung cancer A549 cells(P<0.05);the combination group was superior to the flavonoids from Hedyotis diffusa willd.There were statistically significant differences in the flavonoids from Hedyotis diffusa willd and cisplatin groups(P<0.05).The effect of the combined intervention of the two drugs was simple addition.2.After using drugs to interfere with lung cancer A549 cells for 48 hours,the proportion of G1 phase cells increased in the flavonoids from Hedyotis diffusa willd,and the proportion of cells in S and G2 phase decreased.There was a statistically significant difference compared with the negative control group(P<0.05).The proportion of S phase in the cisplatin group increased and there was a statistically significant difference compared with the negative control group(P<0.05).Compared with the negative control group and the cisplatin group,the proportion of cells in the G1 phase of the combined group increased,compared with the negative control group.Compared with FHD group,the proportion of S phase in the combined group was increased,and the difference was statistically significant(P<0.05).The flavonoids from Hedyotis diffusa willd and cisplatin of Hedyotis diffusa could induce apoptosis of lung cancer A549 cells,and the apoptotic rates of the flavonoids from Hedyotis diffusa willd and cisplatin were(10.870±0.442)% and(16.648±0.307)%,respectively.The ratio of [(1.152±0.275)%] in the control group was statistically significant(P<0.05),and the apoptotic rate of A549 cells in the combined group was [(19.856±0.649)%],which was higher than that of the flavonoids from Hedyotis diffusa willd and cisplatin.There was an increase in the group and the difference was statistically significant.3.Compared with the negative control group,the flavonoids from Hedyotis diffusa willd and cisplatin were significantly inhibiting the expression of p-AKT and Bcl-2 in the lung cancer,and significantly promoted the expression of Caspase-9 and Bax protein.Compared with the negative control group and the single drug intervention group,the synergistic effect of the total flavonoids combined with cisplatin intervention group was significantly enhance,and the difference was statistically significant(P<0.05).Conclusions:1.Both the flavonoids from Hedyotis diffusa willd and cisplatin in the white flowers have the effect of inhibiting the proliferation and promoting apoptosis of the lung cancer A549 cells,and the inhibitory effect of the combination of the two drugs should be significantly enhanced.2.Spreading the flavonoids from Hedyotis diffusa willd can synergy cisplatin,promote the lung cancer A549 cells apoptosis,the mechanism of action is probably by reducing the degree of the expression of p-AKT,increasing the extent of the expression of caspase-9,cut the expression of Bcl-2,enhance the expression of Bax,increase the ratio of Bax/Bcl-2.