Effect of Pulsatilla saponin D on epithelial-mesenchymal transition of colorectal cancer by regulation of Wnt/β-catenin signaling pathway
Objective To investigate the effects of Pulsatilla saponin D(PSD)on the epithelial-mesenchymal transition(EMT)of colorectal cancer,and explore its action mechanisms based on the wingless type MMTV integration site family(Wnt)/β-catenin signaling pathway.Methods Effect of PSD on the cell viability of LoVo cells was determined with the MTT assay.Effect of PSD on the migration of LoVo cells was assessed with the scratch wound healing assay.Effect of PSD on the invasion of LoVo cells was examined with the Transwell invasion assay.Effect of PSD on the malignant transformation of LoVo cells was appraised with the anchorage-independent colony formation assay.Real-time fluorescent quantitative polymerase chain reaction(qRT-PCR)analysis was carried out to detect the effect of PSD on the mRNA levels of EMT-related genes and the genes of Wnt/β-catenin signaling pathway in LoVo cells.Western blotting was performed to measure the effect of PSD on the expression levels of EMT-related proteins and the proteins of Wnt/β-catenin signaling pathway in LoVo cells.Results PSD significantly inhibited the proliferation activity,migration,invasion and colony formation of LoVo cells.Compared with the control group,PSD could reduce the mRNA and protein levels of EMT-related Vimentin and Snail(P＜0.05,0.01),increased the mRNA and protein levels of E-cadherin(P＜0.01).It decreased the mRNA and protein levels of Wnt3a,β-catenin,T-cell factor 4(TCF4),and myelocytomatosis viral oncogene homolog(c-Myc)in Wnt/β-catenin signaling pathway(P＜0.05,0.01),increased the mRNA and protein levels of glycogen synthase kinase-3β(GSK-3β)(P＜0.05,0.01).Furthermore,the inhibition of cell migration and the regulation of the expressions of E-cadherin and vimentin in LoVo cells by PSD were synergized by Wnt/β-catenin signaling pathway inhibitor IWR-1,and antagonized by Wnt/β-catenin signaling pathway activator SKL2001.Conclusion PSD may suppress the EMT of colorectal cancer,potentially by inhibiting the activity of Wnt/β-catenin signaling pathway.
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