Effect of Jinfukang oral liquid on aerobic glycolysis of gefitinib resistant lung adenocarcinoma H1975 cells
Objective It is to investigate the effect of Jinfukang oral liquid(JFK)combined with gefitinib on aerobic glycolysis of lung adenocarcinoma H1975 cells.Methods Firstly,the effects of different concentrations of JFK and gefitinib on the proliferation of H1975 cells were detected by cell counting kit(CCK-8).Then the experiment was divided into blank group,gefitinib group,JFK group and combination group,the glucose uptake and lactate production of the cells in each group were detected by 2-NBDG and lactate test kit,the activities of key rate-limiting enzymes of glycolysis[hexokinase(HK),phosphofructokinase(PFK),pyruvate kinase(PK)]in cells were detected by enzyme activity kit,the protein ex-pressions of hexokinase 2(HK2),P-type phosphofructokinase(PFKP),and M2-type pyruvate kinase(PKM2)in cells were detected by Western blot,and the cellular mitochondrial membrane potential and the level of reactive oxygen species(ROS)were detected by JC-1,DCFH-DA JC-1,DCFH-DA fluorescent probes.Results The growth inhibitory effect of JFK on H1975 cells was enhanced in a time-and dose-dependent manner,and the differences were statistically significant(all P＜0.05);compared with the different concentrations of gefitinib alone for 48 h,the proliferation inhibition rate of H1975 cells after treatment with the corresponding concentration of gefitinib combined with JFK was more higher(all P＜0.05).The differences in the glucose uptake of cells in each treatment group were not significantly different compared with the blank group(P＞0.05),but the cellular lactate production in each treatment group was significantly lower than that in the blank group(P＜0.05),and the combination group was significantly lower than the gefitinib group and JFK group(P＜0.05);the activities of PFK and PK in the gefitinib group,those of HK and PFK in the JFK group,HK,PFK and PK in the combination group were significantly lower than those in the blank group(all P＜0.05),and the activities of HK and PFK in the JFK group and the combination group were significantly lower than those in the gefitinib group(all P＜0.05).The relative expressions of HK2,PFKP and PKM2 proteins in the JFK group and the combination group and the relative ex-pression of PFKP protein in the gefitinib group were significantly lower than those in the blank group(all P＜0.05),and the relative expressions of HK2,PFKP and PKM2 proteins in the JFK group and the combination group were significantly lower than those in the gefitinib group(all P＜0.05).Compared with the blank group,the cellular mitochondrial mem-brane potential was significantly decreased,and the level of ROS was significantly increased in each treatment group,and the decrease and increase were more obvious in the combination group.Conclusion Jinfukang oral liquid can increase the sensitivity of H1975 cells to gefitinib by inhibiting aerobic glycolysis pathway,reducing mitochondrial membrane potential,and promoting the production of ROS.
万方数据
维普
