Inhibitory effect of siRNA specific for EDAG-1 on growth of leukemia cells and the secretion of IL-8 / 肿瘤

ABSTRACT

Objective: To explore the effects of silencing the expression of embryonic develop-associated gene-1 (EDAG-1) by siRNA on the growth of leukemia cell lines and the action mechanisms. Methods: The specific oligonucleotides targeting EDAG-1 were cloned into retroviral vector. The recombinant retroviral vectors were transfected into RetroPack PT67 cell line. The supernatant was collected from the cultured transfected PT67 cells and transfected into HEL cells. The stable transfected HEL cell lines were selected with puromycin. The down-regulation of EDAG-1 expression was determined by RT-PCR. The proliferation of HEL cells was evaluated by MTT assay. The release of IL-8 was analyzed by RT-PCR and ELISA. Results: The recombinant retroviral vectors were successfully constructed. The stable HEL cell line with a persistent knockdown of EDAG-1 gene was obtained by screening. Down-regulation of EDAG-1 expression by siRNA inhibited the proliferation speed of cells and reduced the secretion of IL-8. Conclusion: Silencing the expression of EDAG-1 slowed down the proliferation of leukemia cell ines and decreased the secretion of IL-8, suggesting that EDAG-1 has the potential to become an effective target for leukemia therapy.