Effect of DUSP-1 overexpression on proliferation and invasion of renal cell carcinoma cell line A 498

Objective To construct the pLV-EGFP (2A) Puro-DUSP-1 lentiviral expression vector and package virus and observe the effect of DUSP-1 overexpression on proliferation and invasion of human renal cell carcinoma cell line A498. Methods The target gene obtained from pCMV6-XL5-DUSP-1 was cloned into Plv-EGFP (2A) Puro vector by recombinant DNA technology. The recombinant lentiviral vector pLV-EGFP (2A) Puro-DUSP-1 was co-transfected into 293T cells with packaging plasmids pH1 and pH2 to package virus. The A498 cell line was infected by virus supernatant, and then western blot was used to detect the expression of DUSP-1 protein. The effects of DUSP-1 on proliferation and invasion of A498 cells were analyzed using MTS and Transwell method. Results After successful construction of the recombinant pLV-EGFP (2A) Puro-DUSP-1 lentiviral expression vector, packaging of lentiviral particles and transfection of A498 cell line, the expression of DUSP-1 was significantly upregulated in protein level compared with the empty vector group. MTS assay showed that the absorbance in the recombinant vector group was significantly increased after transfection (P< 0.05) and transwell assay showed that cell invasion numbers in the recombinant vector group were also increased compared with the empty vector group (P< 0.05). Conclusion Recombinant pLV-EGFP (2A) Puro-DUSP-1 lentiviral expression vector is successfully constructed and the lentiviral particles are successfully packaged. The overexpression of DUSP-1 can significantly increase cell proliferation and invasion of A498 cell line.