Mechanism of Ganxi Pill in inhibiting epithelial-mesenchymal transition of HepG2 cells through miR200a/ZEB signaling pathway
Objective To investigate the mechanism by which Ganxi Pill inhibits epithelial-mesenchymal transition(EMT)in HepG2 cells through the microRNA200a/zinc finger E-Box binding homeobox 1(miR200a/ZEB)signaling pathway.Methods HepG2 cells were cultured and treated with Ganxi Pill-medicated serum at different concentrations(10％,15％,and 20％),with a blank control group set up in parallel.The CCK-8 assay was used to assess the proliferation of HepG2 cells,while scratch assay and Transwell assay were conducted to evaluate the horizontal and vertical migration of the cells,respectively.Western blot was employed to check the expressions of ZEB1,ZEB2,E-cadherin,and vimentin.Additionally,RT-PCR was utilized to measure the mRNA expression levels of miR200,ZEB1,ZEB2,E-cadherin,and vimentin.Results Compared with the blank control group,the cell survival rate in all Ganxi Pill groups was significantly reduced,exhibiting clear time-and concentration-dependence(P＜0.05,P＜0.01).The horizontal migration rate and the number of vertically migrating cells significantly decreased in all Ganxi Pill groups(P＜0.05,P＜0.01).Compared with the blank control group,the expression of miR200a increased in the medium-and high-dose Ganxi Pill groups;E-cadherin protein level was elevated in all Ganxi Pill groups,with its mRNA expression increased in the medium-and high-dose groups.Meanwhile,the protein expression of vimentin was reduced across all Ganxi Pill groups,with its mRNA expression decreased in the medium-and high-dose groups.The protein and mRNA expressions of ZEB1 and ZEB2 were reduced in all Ganxi Pill groups(P＜0.05,P＜0.01).Compared with the low-dose group,the medium-and high-dose Ganxi Pill groups exhibited increased protein and mRNA expressions of miR200a and E-cadherin,along with reduced protein and mRNA expressions of vimentin,ZEB1,and ZEB2(P＜0.05,P＜0.01).Compared with the medium-dose group,the high-dose Ganxi Pill group showed further increases in the protein and mRNA expressions of miR-200a and E-cadherin,while the protein and mRNA expressions of vimentin,ZEB1,and ZEB2 decreased(P＜0.05,P＜0.01).Conclusion Ganxi Pill can significantly inhibit the proliferation and migration of HepG2 cells,and its mechanism might be related to the regulation of the miR200a/ZEB signaling pathway,promotion of the E-hadherin expression,inhibition of the vimentin expression,and reversal of EMT.
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