Inhibition of mesangial cell proliferation by antisense oligonucleotides targeting protooncogene mRNA

Abstract

Mesangial cell (MC) proliferation is the hallmark of various forms of renal disease. Many protooncogenes are involved in the signal transduction pathways leading to cell proliferation induced by a wide variety of growth factors. We evaluated the inhibitory effects of antisense oligonucleotides (AS-ODNs) targeting protooncogenes on MC proliferation. The antisense and sense phosphorothioate ODNs of c-myc, c-fos and c-myb were synthesized. Quiescent MCs were stimulated with 20% FCS in the presence of AS-ODN or control sense ODN, which were mixed with liposome. Cell proliferation was determined by counting the number of trypan blue-viable cells. Inhibition of protooncogene mRNA was monitored by reverse transcriptase polymerase chain reaction (RT-PCR) . AS-ODN inhibited MC proliferation in a dose-dependent manner. Inhibitory effects of 10μM of each AS-ODN for MC proliferation were : c-myc 56%, c-myb 45% and c-fos 12%. Sense-ODN had little effect. Any significant morphological change in MC was not noticed with ASODN treatment.
These results indicate that these protooncogenes are involved in the signal transduction pathways mediating MC proliferation. Furthermore, the results suggest a potential role of antisense strategies designed to inhibit protooncogene expression to prevent MC proliferation.