Original Research

Cytotechnology

, Volume 54, Issue 2, pp 97-105

First online:

Influence of glucose starvation on the pathway of death in insect cell line Sl: apoptosis follows autophagy

  • Kaiyu LiuAffiliated withCollege of Life Science, Central China Normal UniversityKey Laboratory of Pesticide & Chemical Biology, Ministry of Education, Central China Normal University Email author 
  • , Qinghua TangAffiliated withCollege of Life Science, Central China Normal University
  • , Cong FuAffiliated withCollege of Life Science, Central China Normal University
  • , Jianxin PengAffiliated withCollege of Life Science, Central China Normal UniversityKey Laboratory of Pesticide & Chemical Biology, Ministry of Education, Central China Normal University
  • , Hong YangAffiliated withCollege of Life Science, Central China Normal UniversityKey Laboratory of Pesticide & Chemical Biology, Ministry of Education, Central China Normal University
  • , Yi LiAffiliated withCollege of Life Science, Central China Normal UniversityKey Laboratory of Pesticide & Chemical Biology, Ministry of Education, Central China Normal University
  • , Huazhu HongAffiliated withCollege of Life Science, Central China Normal UniversityKey Laboratory of Pesticide & Chemical Biology, Ministry of Education, Central China Normal University

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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.

Keywords

Spodoptera litura cell Glucose starvation Apoptosis Autophagy Lysosome Mitochondrion