Repressive effects of RNA interference technique on expression and function of glucocorticoid receptor in human macrophage cell line U937 / 第三军医大学学报

ABSTRACT

Objective To establish a glucocorticoid receptor knockdown model of human macrophage cell line U937 with RNA interference technique. Methods Two RNAi recombinant plasmids (named pSilencer 3.1-GR 1 and pSilencer 3.1-GR 2) targeting to GR gene were constructed. After RNAi recombinant plasmids were transfected into human macrophage cell line U937, the expressions of GR mRNA and GR protein were evaluated with RT-PCR and Western blotting respectively. The transcriptional activation function of GR was evaluated through the detection of relative luciferase activity after dexamethasone treatment. Results Two RNAi recombinant expression plamids were constructed and identified by sequencing. pSilencer 3.1-GR 2 transfection could inhibit not only GR mRNA and protein expressions, but also transcriptional activation function of GR specially; pSilencer 3.1-GR 1 transfection had no significant changes as compared to normal control. Conclusion A glucocorticoid receptor knockdown model has been established successfully, which offers a new method for the further research of GR biological functions.