Synergistic antitumor effect of apigenin combined with trastuzumab on HER2-positive breast cancer
Objective To explore the anti-tumor effect and mechanism of apigenin combined with trastuzumab on human ep-idermal growth factor receptor 2(HER2)positive breast cancer cell lines BT474 and SK-BR-3.Methods The binding ac-tivity of trastuzumab to human breast cancer cells was detected by flow cytometry;Cell counting kit(CCK)-8 was used to detect the growth inhibitory activity of apigenin,trastuzumab monotherapy,and their combination on BT474 and SK-BR-3 cells;EdU staining method was used to detect the effects of apigenin,trastuzumab monotherapy,and their combination on DNA replication in BT474 and SK-BR-3 cells;Protein blotting was used to detect the effects of apigenin,trastuzumab monotherapy,and their combination on the expression levels of epidermal growth factor receptor(EGFR),HER2,HER3,Akt,and their respective phosphorylated proteins in BT474 and SK-BR-3 cells.The experiment was divided into apigenin group,trastuzumab group,apigenin+trastuzumab combination group,control group,and blank control group.Results The results of the growth inhibition experiment showed that the IC50 values of apigenin inhibiting the growth of BT474 and SK-BR-3 cells were 25.70 and 29.34 μmol/L,respectively.The EdU staining experiment results showed that after different concentrations of apigenin(16,32,64 μmol/L)were applied to SK-BR-3 and BT474 cells,and there were statistically significant differences compared to the control group,with F values of 99.25 and 309.10,both P＜0.001.After treating SK-BR-3 and BT474 cells with a combination of apigenin and trastuzumab,there was a statistically signifi-cant difference in the positive rate of EdU compared to the apigenin group,with F values of 145.60 and 110.10,respec-tively,all P＜0.001;There was a statistically significant difference in the positive rate of EdU compared to the trastuzum-ab group,with F values of 48.38 and 66.46,respectively,both P＜0.001.The protein blot results showed that the rela-tive expression levels of p-EGFR protein in the apigenin group,trastuzumab group,and apigenin+trastuzumab combina-tion group were(81.75±1.08)％,(45.84±1.71)％and(17.06±1.06)％,F=2.093.00,P＜0.001.The relative ex-pression levels of p-HER2 protein were(55.09±6.23)％,(41.33±2.48)％and(8.87±0.20)％,F=604.60,P＜0.001.The relative expression levels of p-HER3 protein were(67.78±1.94)％,(17.89±0.84)％and(12.06±0.81)％,F=2 749.00,P＜0.001.The relative expression levels of p-Akt protein were(46.85±13.96)％,(42.35±2.71)％and(23.41±0.24)％,F=85.73,P＜0.001.Conclusion Apigenin and trastuzumab can synergistically inhibit the proliferation of HER2 positive breast cancer cells,and the mechanism may be related to the inhibition of Akt signaling pathway.
breast cancertrastuzumabcelery extracthuman epidermal growth factor receptor 2
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