Objective To explore the effect and mechanism of GATA zinc finger domain containing 1(GATAD1)on the migration of colon cancer cells.Methods The GATAD1 interference plasmid SiRNA-GATAD1(Si-GATAD1)was constructed using the restriction endonuclease ligation method.Normal human colon epithelial cells NCM460 and colon cancer cells Caco-2,HCT-116,HCT-8,HT-29,and RKO were cultured separately.The relative expression levels of GATAD1 protein in each group were detected by Western blot.Colon cancer cells HCT-116 and RKO with relatively high GATAD1 expression levels were selected for transfection of GATAD1 interference plasmid Si-GATAD1,and colon cancer cells Caco-2 with the lowest GATAD1 expression levels were selected for transfection of overexpressing pMZ-GATAD1 plasmid.HCT-116 and RKO cells were randomly divided into the negative control group(NC group)and the Si-GATAD1 plasmid transfection group(Si-GATAD1 group).Caco-2 cells were randomly divided into the NC group and the overexpressing pMZ-GATAD1 plasmid transfection group(pMZ-GATAD1 group).Si-GATAD1 plasmids were transfected into HCT-116 and RKO cells,respectively,in the Si-GATAD1 group.The overexpressing pMZ-GATAD1 plasmids were transfected into Caco-2 cells in the pMZ-GATAD1 group.The empty pML-MO3 plasmids were transfected into HCT-116,RKO and Caco-2 cells in the NC group.The cell scratch assay was used to detect the scratch healing rate of cells in each group.Western blot was used to detect the relative expression levels of phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway related proteins PI3K,Akt,phosphorylated protein kinase B(p-Akt),and Cyclin D1(CCND1)in each group.Results The relative expression levels of GATAD1 protein in colon cancer cells HCT-8,HCT-116,RKO,HT-29,and Caco-2 were significantly higher than those in normal human colon epithelial cells NCM460(P＜0.05).The relative expression levels of GATAD1 protein in colon cancer cells HCT-116 and RKO were significantly higher than those in HCT-8,HCT-29,and Caco-2 cells(P＜0.05),while the relative expression level of GATAD1 protein in Caco-2 cells was significantly lower than that in HCT-8 and HCT-29 cells(P＜0.05).The rela-tive expression levels of GATAD1 protein in HCT-116 and RKO cells in the Si-GATAD1 group were significantly lower than those in the NC group(P＜0.05),while the relative expression level of GATAD1 protein in Caco-2 cells in the pMZ-GATAD1 group was significantly higher than that in the NC group(P＜0.05).After 12 and 24 hours of cultivation,the scratch healing rates of HCT-116 and RKO cells in the Si-GATAD1 group were significantly lower than those in the NC group(P＜0.01),while the scratch healing rates of Caco-2 cells in the pMZ-GATAD1 group were significantly higher than those in the NC group(P＜0.01).The relative expression levels of PI3K,p-Akt,and CCND1 proteins in RKO and HCT-116 cells in the Si-GATAD1 group were significantly lower than those in the NC group(P＜0.05),while the relative expression levels of PI3K,p-Akt,and CCND1 proteins in Caco-2 cells in the pMZ-GATAD1 group were significantly higher than those in the NC group(P＜0.05).There was no statistically significant difference in the relative expression of Akt protein between the Si-GATAD1 group(RKO and HCT-116 cells),pMZ-GATAD1 group(Caco-2 cells)and the NC group(P＞0.05).Conclusion GATAD1 is highly expressed in colon cancer cells and promotes their migration by regulating the PI3K/Akt signaling pathway.
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