ABSTRACT
Objective:To investigate the influence of cereblon(CRBN)on the inhibitory effect of thalidomide on the migration of human lung adenocarcinoma A549 cells and related mechanism. Methods: The effect of thalidomide on the A549 cell proliferation was assayed by the MTS method. CRBN in A549 cells was knocked down(A549CRBN cells) with RNA interference and lentivirus infection. Transwell assay was performed to detect the effect of thalidomide on the cell migration of A549 and A549CRBN cells. RT-qPCR assay was performed to detect the gene expression of N-cadherin and vimentin. Results: The thalidomide at the 1, 10, 50 and 100 μmol/L concentration did not affect the proliferation of A549 cells, while the 100 μmol/L thalidomide could significantly inhibit the migration of A549 cells. After the CRBN gene knockdown the migration ability of A549 cells was significantly decreased(P<0.01), and the gene expression of N-cadherin and vimentin was also significantly reduced in the A549CRBN cells. The inhibitory effect of thalidomide on the cell migration in the A549 cells disappeared after CRBN gene knockdown. Thalidomide could inhibit the gene expression of N-cadherin and vimentin in a CRBN-dependent manner. Conclusion: Thalidomide could inhibit the migration of A549 cells and the expression of the migration-related genes via CRBN.
ABSTRACT
Objective: To establish a molecular platform for screening thalidomide derivatives with potential high anti-tumor activities. Methods: Human cereblonvCRBN) and Ikaros family zinc finger protein l(IKZF1) cDNA were amplified and cloned into pXJ40-myc and pcDNA3-FLAG vectors respectively. Then the two constructs were transfected into 293T cells and the anti-tumor activities of thalidomide and its derivatives were reflected by their effects on the binding between CRBN and IKZF1 proteins. Results: Both CRBN and IKZF1 were successfully expressed in 293T cells. Thalidomide and two of its derivatives were found to enhance the interaction between CRBN and IKZF1 significantly. Conclusion: A molecular platform was successfully established for screening thalidomide derivatives with potential high anti-tumor activities and two thalidomide derivatives could potentiate the binding between CRBN and IKZF1. suggesting that they possess potential anti-tumor activities.