Influence of glucose starvation on the pathway of death in insect cell line Sl: apoptosis follows autophagy
Influence of glucose starvation on the pathway of death in insect cell line Sl: apoptosis follows autophagy
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葡萄糖饥饿对昆虫细胞系Sl死亡途径的影响：细胞凋亡跟随自噬
DOI：
10.1007/s10616-007-9080-5
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发表时间：
2007-06-01
期刊：
影响因子：
2.2
通讯作者：
Hong, Huazhu
中科院分区：
文献类型：
--
作者：
Liu, Kaiyu;Tang, Qinghua;Hong, Huazhu
自噬与细胞凋亡之间的关系尚未明确阐明。在这里，我们报道了在经历葡萄糖饥饿的昆虫斜纹夜蛾细胞（Sl）中自噬后发生细胞凋亡。 Sl 细胞已适应补充有葡萄糖 (1.0 g/l) 和 5% 胎牛血清 (FBS) 的 Leibovitz-15 培养基，用于哺乳动物细胞培养。如果在含5% FBS的L-15培养基中不添加葡萄糖（1 g/l）或谷氨酰胺（1.6 g/l），Sl细胞开始形成许多空泡，并且这些空泡在细胞质中逐渐增大，这就是自噬空泡。然而，这些大液泡在葡萄糖饥饿 48 小时后开始逐渐消失，并伴随着显着的细胞凋亡，没有凋亡小体，这通过 DNA 断裂和 caspase-3 样激活来证明。在葡萄糖饥饿期间，S1细胞ATP浓度逐渐下降。有趣的是，如果在培养物严重饥饿48小时或更长时间后，将不含葡萄糖的条件L-15培养基替换为补充有葡萄糖或谷氨酰胺的新鲜L-15培养基，则开始形成凋亡小体。这些数据表明，细胞 ATP 的部分耗尽引发了葡萄糖饥饿的 Sl 细胞自噬后的细胞凋亡，并且凋亡小体的形成需要比 DNA 断裂和 caspase-3 样活性更高水平的 ATP。此外，饥饿后期（超过48 h）Sl细胞自噬空泡的消失、中性红阴性染色、吖啶橙绿染色以及酸性磷酸酶活性扩散表明溶酶体功能障碍更可能参与细胞凋亡。放线菌素 D 诱导的细胞凋亡受到部分抑制，而环孢菌素 A 阻断线粒体通透性转变（MPT）孔的开放，部分抑制葡萄糖饥饿的 Sl 细胞的凋亡，这表明葡萄糖饥饿诱导的细胞凋亡途径似乎与放线菌素 D 诱导的细胞凋亡和 MPT 孔的开放不同。 线粒体可能分别参与由葡萄糖饥饿引发的细胞凋亡。
The relation between autophagy and apoptosis has not been clearly elucidated. Here, we reported that apoptosis followed autophagy in insectSpodoptera lituracells (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.