Influence of glucose starvation on the pathway of death in insect cell line Sl: apoptosis follows autophagy |
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Authors: | Kaiyu Liu Qinghua Tang Cong Fu Jianxin Peng Hong Yang Yi Li Huazhu Hong |
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Institution: | (1) College of Life Science, Central China Normal University, Wuhan, 430079, China;(2) Key Laboratory of Pesticide & Chemical Biology, Ministry of Education, Central China Normal University, Wuhan, 430079, China |
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Abstract: | The relation between autophagy and apoptosis has not been clearly elucidated. Here, we reported that apoptosis followed autophagy
in insect Spodoptera litura cells (Sl) undergoing glucose starvation. Sl cells have been adapted to Leibovitz-15 medium supplemented with glucose (1.0 g/l)
and 5% fetal bovine serum (FBS), used for mammalian cell cultures. If glucose (1 g/l) or glutamine (1.6 g/l) had not been
supplemented in L-15 medium with 5% FBS, Sl cells began to form many vacuoles and these vacuoles gradually enlarged in the
cytoplasm, which were autophagic vacuoles. However, these large vacuoles began to disappear gradually after 48 h of glucose
starvation, accompanied with remarkable apoptosis without apoptotic bodies, which was demonstrated by DNA fragmentation and
activation of caspase-3-like. During glucose starvation, Sl cell ATP concentrations gradually decreased. Interestingly, if
the conditioned L-15 medium without glucose was replaced with fresh L-15 medium supplemented with glucose or glutamine after
the cultures had been starved seriously for 48 h or longer, the formation of apoptotic bodies was initiated. These data suggested
that the partial depletion of cell ATP triggered apoptosis following autophagy in glucose-starved Sl cells and the formation
of apoptotic bodies required higher level of ATP than DNA fragmentation and activation of caspase-3-like activity. Additionally,
the disappearance of autophagic vacuoles, negative staining of neutral red, green staining of acridine orange and diffusion
of acid phosphatase activity in Sl cells at the late stage of starvation (over 48 h) suggested that the dysfunction of lysosome
was more likely to involve in apoptosis. The facts that Actinomycin D-induced apoptosis was partially inhibited and cyclosporin
A, blocking the opening of mitochondrial permeability transition (MPT) pores, inhibited partially apoptosis in glucose-starved
Sl cells, suggested the pathway of glucose starvation-induced apoptosis seemed to be different from that induced by actinomycin
D and the opening of MPT pores on mitochondria probably involved in apoptosis triggered by glucose starvation, respectively. |
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Keywords: | Spodoptera litura cell Glucose starvation Apoptosis Autophagy Lysosome Mitochondrion |
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