Inhibitory effects of Biejiajian Pill on subcutaneous tumor of MHCC-97H hepatoma cells based on EGFR/MAPK/ERK signaling pathway
Objective To investigate the inhibitory effects and their mechanism of Biejiajian Pill(BJJP)on the subcutaneous tumor of MHCC-97H hepatoma cells based on the signaling pathway of epidermal growth factor receptor(EGFR)/mitogen-activated protein kinase(MAPK)/extracellular signal-regulated kinase(ERK).Methods A total of 30 male BLAB/c nude mice were used to establish a subcutaneous tumor model of MHCC-97H hepatoma cells.After successful modeling,they were randomized into model group,low-,medium-,and high-dose BJJP groups,and western medicine group,with six mice in each group.Low-,medium-,and high-dose BJJP groups were treated with concentrated BJJP 0.55 g/kg,1.1 g/kg,and 2.2 g/kg,respectively,twice a day;western medicine group was administered Lenvatinib 4 mg/kg combined with Gefitinib 80 mg/kg,5 days a week;model group was given the equal volume of normal saline;twice a day.The intervention was carried out by gavage for 14 days successively.The general condition of mice was observed,and the tumor inhibition rate of each group was calculated.HE staining was used to observe pathological and morphological changes;RT-qPCR was used to determine the mRNA expression levels of EGFR,mitogen-activated protein kinase kinase(MEK),ERK1,and ERK2 in the tumor tissue;Western blot was used to examine the relative expression levels of EGFR,phosphorylated EGFR(p-EGFR),MEK,phosphorylated MEK(p-MEK),ERK1,ERK2,phosphorylated ERK1/2(p-ERK1/2),matrix metalloproteinase-1(MMP-1),cell cycle protein D1(Cyclin D1),N-cadherin,and E-cadherin.Results Compared with model group,the condition of spirit,reactions,and intake of food and water of mice in low-,medium-,and high-dose BJJP groups and western medicine group were improved significantly.Compared with day 0,the body weight of mice in each group was significantly lower on the 14th day,with the most obvious reduction in western medicine group(P＜0.01).Compared with model and low-dose BJJP groups,the tumor mass of mice in medium-and high-dose BJJP groups and western medicine group was significantly reduced(P＜0.05,P＜0.01).The tumor inhibition rates of low-,medium-,and high-dose BJJP groups and western medicine group were 20%,47.73%,55.91%,and 75.45%,respectively.Compared with model group,the tumor cells in the low-,medium-,and high-dose BJJP groups and western medicine group were loosely arranged and the boundaries were blurred,and the cell nuclei were condensed and ruptured,especially in western medicine group.Compared with model group,the mRNA expression levels of EGFR,MEK,ERK1,and ERK2 in low-,medium-,and high-dose BJJP groups and western medicine group significantly decreased(P＜0.01);compared with low-dose BJJP group,the mRNA expression levels of EGFR,MEK,ERK1,and ERK2 in high-dose BJJP and western medicine groups were significantly reduced(P＜0.01),and that of ERK1 in medium-dose BJJP group was significantly lower(P＜0.01);compared with medium-dose BJJP group,the mRNA expression levels of EGFR and ERK2 in western medicine group decreased(P＜0.05,P＜0.01).Compared with model group,medium-and high-dose BJJP groups and western medicine group showed a decrease in the relative protein expression levels of p-EGFR/EGFR,p-MEK/MEK,p-ERK1/ERK1,p-ERK2/ERK2,MMP-1,Cyclin D1,and N-cadherin(P＜0.05,P＜0.01),and a significant increase in the relative protein expression level of E-cadherin(P＜0.01).Compared with high-dose BJJP group,the relative protein expression levels of p-EGFR/EGFR,p-ERK1/ERK1,MMP-1,Cyclin D1,and N-cadherin in western medicine group were significantly reduced(P＜0.01),while that of E-cadherin was elevated(P＜0.05).Conclusion BJJP may downregulate the protein expressions of MMP-1,Cyclin D1,and N-cadherin and upregulate that of E-cadherin by inhibiting EGFR/MAPK/ERK signaling pathway activation,thereby exerting significant inhibitory effects on the subcutaneous tumor of MHCC-97H hepatoma cells.
Biejiajian Pillepidermal growth factor receptormitogen-activated protein kinaseextracellular signal-regulated kinaseprimary liver cancercirculating blood and eliminating blood stasis
万方数据
维普
