INHIBITION OF MESANGIAL CELL-PROLIFERATION BY E2F DECOY OLIGODEOXYNUCLEOTIDE IN-VITRO AND IN-VIVO

The transcription factor E2F coordinately activates several cell cycle -regulatory genes, We attempted to inhibit the proliferation of mesang ial cells in vitro and in vivo by inhibiting E2F activity using a 25-b p decoy oligodeoxynucleotide that contained consensus E2F binding site sequence (E2F-decoy) as a competitive inhibitor. The decoy's effect o n human mesangial cell proliferation was evaluated by [H-3]thymidine i ncorporation. The E2F decoy inhibited proliferation in a concentration -dependent manner, whereas a mismatch control oligodeoxynucleotide had little effect. Electrophoretic mobility shift assays demonstrated tha t the decoy's inhibitory effect was due to the binding of the decoy ol igodeoxynucleotide to E2F, The effect of the E2F decor was then tested in a rat anti-Thy 1.1 glomerulonephritis model. The E2F decoy oligode oxynucleotide was introduced into the left kidney 36 h after the induc tion of glomerulonephritis. The administration of E2F decoy suppressed the proliferation of mesangial cells by 71%. Furthermore, treatment w ith the E2F decoy inhibited the glomerular expression of proliferating cell nuclear antigen at the protein level as well as the mRNA level. These findings indicate that decoy oligonucleotides can suppress the a ctivity of the transcription factor E2F, and may thus have a potential in treating glomerulonephritis.