Abstract
Downregulation of microRNA-31 has been linked with enhanced stress resistance, while its overexpression leads to cell death. In this study, we found mediatory role of microRNA-31 in γ-radiation-induced apoptosis in a model insect cell line Sf9 carrying well-conserved apoptotic machinery. Mature microRNA-31 is perfectly conserved amongst insects; hence we used biotinylated probes designed from Bombyx mori sequence for its successful detection in Sf9 cells. Target identification using Bombyx mori 3'UTRs predicted miR-31's potential role in Lepidopteran apoptosis, which prompted us to investigate alterations in its expression during radiation-induced cell death. We found significant overexpression of Sf-miR-31 following lethal dose (1,000Gy-3,000Gy) irradiation. Its mediatory role was finally confirmed as antisense-microRNA-31 could successfully inhibit radiation-induced cytochrome-c release, caspase-3 activation and apoptosis. While Bax/Bcl-2 expression remained unchanged, lethal radiation doses induced Bim overexpression and direct Bim-Bax interaction (co-immunoprecipitation) which is not yet unequivocally demonstrated during apoptosis. Quite important, these events were found to be dependent on radiation-induced miR-31 overexpression, as antisense-miR-31 inhibited both the responses and resulted in significant inhibition of cell death. Pro-apoptotic role of miR-31 was further confirmed when miR-31 mimic induced apoptosis involving similar Bim/Bax alterations. Therefore, our study reveals an important mediatory role of miR-31 in radiation-induced cell death.