Up-regulation of miR-378b inhibits epithelial-mesenchymal transition,invasion and migration of thyroid cancer cells by inhibiting ALX4 methylation
Objective To investigate the ability of miR-378b to regulate epithelial-mesenchymal transition,invasion and migration of thyroid cancer cells through ALX4.Methods The methylation status of ALX4 gene in thyroid cancer cells was detected by MSP method,the thyroid cancer cells were treated with the demethylation drug 5-aza-dc,and the relationship between ALX4 expression and promoter methylation was investigated by MSP,RT-PCR and Western blot.miR-378b mimics/inhibitors were transfected into thyroid cancer cells,and the effect of miR-378b on ALX4 methylation was detected by MSP.The effects of miR-378b on the expression of ALX4 and epithelial-mesenchymal factors HMGA1,N-cadherin,and Vimentin were detected by RT-PCR and Western blot,and the invasion and migration of thyroid cancer cells were detected by TranswellTM.The targeting relationship between miR-378b and ALX4 was determined by double luciferase activity analysis experiments.After pcDNA3.1-ALX4 plasmid was overexpressed,the effect of ALX4 on cell epithelial-mesenchymal transition and its ability to invade and migrate was examined.Results Compared with normal thyroid cells,ALX4 gene was methylated in thyroid cancer cells,and the expression of ALX4 was suppressed due to the occurrence of methylation.MiR-378b mimics inhibited the occurrence of ALX4 methylation,promoted the expression of ALX4,inhibited the expression of epithelial-mesenchymal transition-related factors HMGA1,N-cadherin and Vimentin,and inhibited cell invasion and migration ability.MiR-378b targets the 3'untranslated region(3'UTR)of the ALX4 gene.Overexpression of pcDNA3.1-ALX4 inhibited the expression of HMGA1,N-cadherin and Vimentin,and inhibited the invasion and migration ability of the cells.Conclusion miR-378b inhibits the epithelial-mesenchymal transition and cell invasion and migration of thyroid cancer cells via inhibiting the occurrence of ALX4 methylation.
thyroid cancer cellsmiR-378bALX4 methylationcell epithelial-mesenchymal transitioninvasion and migration
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