SOX2 And Wnt/β-catenin Signaling Pathway On Epithelial Mesenchymal Transformation Of Colorectal Cancer Cell Lines SW620

Objectives To explore the mechanism and relationship between SOX2 and Wnt/β-catenin signaling pathway in the occurrence and development of epithelial-mesenchymal transition by restraining the expression of SOX2 m RNAofcolorectal cancer cell line SW620 through lentiviral-mediated plasmid interference technique.Methods 1 After screening the resistant strains of colorectal cancer cell line SW620 trasfeced SOX sh RNA by lentiviral-mediated plasmid interference technique,the cell morphological changes of the gene interference group and the control group were observed by the inverted phase contrast microscope. 2 The migration ability was detected by scratch experiment in the both group. 3 The expression of m RNA or protein of SOX2, β-catenin, E-cadherin and Vimentin were detected respectively by RT-PCR or Western Blot.Results 1 Cell morphology: Comparing with in the control group, the morphologic of cell line SW620 in the interference group, after SOX2 sh RNA silence, changedfrom long spindle to round or ovoid, pseudopodia decreased and cell density increased. 2 The scratch test: After SOX2 sh RNA silence, cell migration ability of cell line SW620 decreased in the interference group. 3 RT-PCR: The m RNA relative quantity of SOX2, β-catenin and Vimentin in the gene interference group were 0.25±0.04, 0.20±0.03, and 0.19±0.03, which were significantly lower than that in the control group(P <0.05). While the m RNA relative quantity of E-cadherin was 0.87±0.04, which was significantly higher than that in the control group(P <0.05). The correlation analysis showed the m RNA expression of SOX2 and β-catenin were positively correlated(R = 0.910, P =0.012), the m RNA expression of SOX2 and Vimentin were positively correlated(R =0.829, P=0.042), while the m RNA expression of SOX2 and E-cadherin were negatively correlated(R=-0.841, P =0.036); the m RNA expression of β-catenin and vimentin were positively correlated(R= 0.886, P =0.019), while the m RNA expression of β-catenin and E-cadherin were also negatively correlated(R=-0.899, P=0.015). 4 Westen blot: In the gene interference group, the protein relative quantity of SOX2, β-catenin and Vimentin were 0.33±0.02, 0.18±0.02 and 0.29±0.02, which were significantly decreased comparing with that in the control group(P<0.05). While the protein relative quantity of E-cadherinprotein was 0.78±0.02,which was significantly higher than that in the control group(P<0.05). Correlation analysis showed that the protein expression of SOX2 and β-catenin were positively correlated(R = 0.841, P=0.036), the protein expression of SOX2 and vimentin were positive correlated(R = 0.829, P=0.042), while the protein expression of SOX2 and E-cadherin were negative correlated(R =-0.928, P=0.008); the protein expression of β-catenin and E-cadherin were negatively correlated(R=-0.870, P =0.024).Conclusions SOX2 may enhance the invasion and migration of colorectal cancer cell line SW620 by promoting epithelial-mesenchymal transition coordinating Wnt / β-catenin signaling pathway.