Abstract
Mitochondria have been shown to play an important role in apoptosis using mammalian cell lines. However, this seems not to be the case in Drosophila, an insect model organism; thus more in-depth studies of insect cell apoptosis are necessary. In the present study, mitochondrial involvement during azadirachtin- and camptothecin-induced apoptosis in Spodoptera frugiperda Sf9 cells (isolated from Spodoptera frugiperda pupal ovarian tissue) was investigated. The results showed that both azadirachtin and camptothecin could induce apoptosis in Sf9 cells. Reactive oxygen species (ROS) generation, activation of mitochondrial permeability transition pores (MPTPs) and loss of mitochondrial membrane potential (MMP) were observed very early during apoptosis and were followed subsequently by the release of cytochrome-c from the mitochondria. Furthermore, the results also revealed that the opening of MPTPs and the loss of MMP induced by azadirachtin could be significantly inhibited by the permeability transition pore (PTP) inhibitor cyclosporin A (CsA), which was used to identify the key role of mitochondria in the apoptosis of Sf9 cells. However, in camptothecin-treated Sf9 cells, CsA could not suppress the opening of MPTPs and the loss of MMP when apoptosis was induced. The data from caspase-3 and caspase-9 activity assays and detection of apoptosis by morphological observation and flow cytometry also uncovered the different effect of CsA on the two botanical apoptosis inducers. Although different mechanisms of apoptosis induction exist, our study revealed that mitochondria play a crucial role in insect cell line apoptosis.
Highlights
Apoptosis is an evolutionarily conserved form of programmed cell death (PCD) that can be induced by particular endogenous and exogenous factors, such as toxins, hormones, growth factors, nitric oxide, cytokines, heat, irradiation, nutrient deprivation, viral infection, hypoxia [1] and increased intracellular calcium concentration [2]
The apoptotic DNA fragmentation evident from the formation of a typical DNA ladder was observed at 12 h and 48 h post treatment after captothecinand azadirachtin-treatement, which further confirmed the induction of apoptosis
The present study aims to further investigate the mechanism of azadirachtininduced apoptosis in Lepidopteran cell lines and the role of mitochondrial permeability transition pores (MPTPs) in apoptosis of insect cell line
Summary
Apoptosis is an evolutionarily conserved form of programmed cell death (PCD) that can be induced by particular endogenous and exogenous factors, such as toxins, hormones, growth factors, nitric oxide, cytokines, heat, irradiation, nutrient deprivation, viral infection, hypoxia [1] and increased intracellular calcium concentration [2]. It is a basic dynamic process, which is essential to eliminate unwanted or abnormal cells and plays an important role in the stability of the internal environment and the development of multicellular organisms [3,4]. There is no confirmation that the mechanism of apoptosis induced by camptothecin in insect cells is the same as that in mammalian cells
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