鱼腥草总黄酮对肝癌细胞耐药性的逆转作用及机制研究OA

Objective:To investigate the effects of total flavonoids of Houttuynia cordata on Bel-7402/ADM cells and the mech-anism of possible reversal action of multidrug resistance(MDR)in hepatocellular carcinoma by observing the effects of total flavo-noids of Houttuynia cordata on the expression of MDR cells of human hepatocellular carcinoma,Bel-7402/ADM,and P-glycoprotein(P-gp).Methods:CCK-8 assay was used to detect drug resistance and inhibition rate of hepatocellular carcinoma cells;The experiment was divided into the parental group(Bel-7402),the parental adriamycin(ADM)group(Bel-7402+ADM),the drug-resistant group(Bel-7402/ADM),the drug-resistant ADM group(Bel-7402/ADM+ADM),the total flavo-noids of Houttuynia cordata group(Bel-7402/ADM+ADM+EAT),and the positive control group(Bel-7402/ADM+ADM+VRP),and the amount of ADM accumulation in the cells of each group after the treatment was detected by flow cytometry and fluores-cence observation;and the experiments were divided into the parental group(Bel-7402),the drug resistant group(Bel-7402/ADM),the total flavonoid of Houttuynia cordata group(Bel-7402/ADM+EAT),and the positive control group(Bel-7402/ADM+VRP),and the expression of P-gp protein in the cells of each group was detected by flow cytometry and immunohistochemistry.Results:ADM inhibited the proliferation of both Bel-7402 cells and Bel-7402/ADM cells,but the effect on the proliferation of Bel-7402 cells was larger;The fluorescence results showed that there was no fluorescence in the parental group and a small amount of fluorescence in the drug-resistant group;the fluorescence intensity of the parental ADM group and the drug-resistant ADM group increased and the drug-resistant ADM group was stronger than the parental ADM group;The fluorescence intensity of the total flavonoids of Houttuynia cordata group and the positive control group was significantly enhanced compared to that of the parental,the drug-resistant,and the parental ADM groups,and slightly weakened compared to the drug-resistant ADM group;The flow cytometry results showed that the fluorescence intensity of ADM in each group was drug-resistant ADM group>positive control group>parental ADM group>total flavonoids of Houttuynia cordata group>drug-resistant group>parental group(P<0.05);The flow cytometry results showed that there was no statistically significant difference in the mean fluorescence intensity(MFI)of P-gp between the parental group and the drug-resistant group(P=0.671),the total flavonoid of Houttuynia cordata group and the positive control group were significantly lower than that of the parental group and the drug-resistant group(P<0.001),and there was no statistically significant difference between the total flavonoid of Houttuynia cordata group and the posi-tive control group(P=0.367);The immunohistochemistry results showed that the positivity rate of the P-gp protein in the paren-tal group was(15.34±2.15)%,compared to that of the drug-resistant group(99.94±0.02)%,the total flavonoid of Houttuynia cordata group(10.45±0.73)%,and the positive control group(1.08±0.13)%,the differences were all statistically significant(P<0.001).Conclusion:The total flavonoids of Houttuynia cordata can inhibit the proliferation of Bel-7402/ADM cells and increase intracellular ADM accumulation,thus may enhance the antitumor effect of ADM and reverse the MDR of hepatocellular carcino-ma,and the mechanism may be related to the down-regulation of the expression of P-gp.