| Objective: To investigate the effect of Forkhead BoxG1(FOXG1)on the epithelial-mesenchymal transition(EMT)of colorectal cancer(CRC)cell lines.Methods:FOXG1 lentiviral interference(sh RNA)plasmid and expression plasmid were constructed.Western blotting was used to analyze the expression of FOXG1 protein in five CRC cells lines,among which were RKO,SW480,SW620,LoVo and DLD-1.The shRNA fragment of FOXG1(sh FOXG1)was designed and synthesized.Recombinant plasmids were obtained with the aid of DNA recombination technique.Double digestion and sequencing were used to identify the recombinant plasmids,and then lentivirus packaging,purification and stable transfection were carried out.Additionally,stable CRC cell lines were screened out.The changes of FOXG1 knockdown and overexpression efficiency,E-cadherin,Vimentin,Fibronectin,Snail,Twist mRNA and protein were investigated respectively by Western blotting and qRT-PCR analysis.Furthermore,the changes of cell morphology after knockdown and cell migration ability were evaluated respectively with optical microscopy,scratch test and Transwell assay.Results:FOXG1 had the highest protein expression in RKO and the lowest in DLD-1 among the five CRC cells.Compared with those of the control group,the cell morphology in FOXG1 knockdown RKO group was changed from spindle into round or polygonal shape,cell polarization wasenhanced and tight junction assembly was accclerated while cell migration distance was noticeably decreased.Moreover,the number of cells invaded and migrated through chambers was significantly reduced.Among these key factors of EMT,the expression of E-cadherin was increased while the expressions of Vimentin,Fibronectin,Snail and Twist were decreased.The opposite was the case in the overexpressed FOXG1 group.Conclusion: The overexpression of FOXG1 in CRC promoted the invasion and metastasis of CRC cells and played a crucial role in regulating the EMT.Thus,FOXG1 might be a novel therapeutic target in CRC treatment. |
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