Responses of two insect cell lines to starvation: Autophagy prevents them from undergoing apoptosis and necrosis, respectively

Abstract

The responses of insect cells to starvation and the characteristics of cell death after the depletion of nutrients remain largely unknown. In the present study, we investigated autophagy, apoptosis and necrosis in two Lepidoptera insect cell lines in response to amino acid starvation. Our data demonstrated that starvation induced a significant increase in autophagy in Spodoptera litura SL-ZSU-1 cells, and cell apoptosis followed autophagy after starvation of more than 48h. However, at an early stage of starvation, inhibition of autophagy with 3-MA rapidly triggered apoptosis of SL-ZSU-1 cells, suggesting autophagy inhibits cell apoptosis. By contrast, Bombyx mori SPC Bm36 cells died by a non-apoptotic pathway if the starvation was prolonged for more than 48h. At the early stage of starvation, inhibition of autophagy with 3-MA did not trigger apoptosis in Bm36 cells, but resulted in necrotic-like cell death. Under starvation pressure, autophagy in SL-ZSU-1 cells was much more active than in Bm36 cells. The activity of caspase-9-like in apoptotic SL-ZSU-1 cells also was much higher than in apoptotic Bm36 cells. RT-PCR analyses revealed that transcriptional levels of saposin-like (Bm109) and Atg6 were undetectable in Bm36 cells, but expression level of saposin-like in SL-ZSU-1 was high. Expression of Atg6 in SL-ZSU-1 cells was not analyzed because its sequence was unknown. These data indicate that autophagy prevents Lepidoptera insect cells from death at an early stage of starvation, but prolonged starvation results in cell death. The pathways of cell death might be dependent on the abundance of caspase-9-like, saposin-like and Atg6.