College of Bioscience and Biotechnology, Insect Resource Research Center for Engineering and Technology of Liaoning Province, Shenyang Agricultural University, Shenyang, China.
USDA/Agricultural Research Service, Biological Control of Insects Research Laboratory, Columbia, Missouri, USA.
Arch Insect Biochem Physiol. 2021 Nov;108(3):e21844. doi: 10.1002/arch.21844. Epub 2021 Sep 13.
Animals maintain homeostasis of cell numbers, constantly creating new cells and eliminating others. Programmed cell death, apoptosis, is a mechanism of cell elimination and it acts in many aspects of animal biology. Drawing on the biomedical background, several signals launch the apoptosis mechanisms, including prostaglandins (PGs). Based on this information, we posed the hypothesis that PGs similarly induce apoptosis in insect cell lines. We used three Spodoptera frugiperda cell lines, including two newly established, BCIRL-SfNS-0518B-YL derived from the central nervous system and BCIRL-Sf4FB-0614-SGS derived from fat body, and the commercially available Sf9 cells. Using a kinetic apoptosis kit, we found treating SfNS cells for 18 h with 15 or 20 μM PGA led to decreases in cell numbers, coupled with increased numbers of apoptotic and dead cells. Similar exposures to 10 μM PGA (24 h) led to substantial increases in apoptotic cells, confirmed by a terminal deoxynucleotidyl transferase dUTP nick end labeling assay on a flow cytometer. The influence of PGA treatments increased with dosage, as we recorded about 20% apoptosis at 24 h post-PGA treatments (10 μM) and about 34% apoptosis at 24 h post-30 μM treatments. PGA treatments led to 10- to 30-fold increases in messenger RNAs (mRNAs) encoding apoptosis-specific caspases-1, -2, -3, and -5 at 12 h and 40- to 60-fold increases in mRNAs encoding caspases-1 and -2, 10-fold increases for caspases-3 and -5 at 24 h. These findings strongly support our hypothesis that PGs induce apoptosis in an insect cell line and confirm an additional PG action in insect biology.
动物维持细胞数量的内稳态,不断产生新细胞并清除其他细胞。程序性细胞死亡,即细胞凋亡,是细胞清除的一种机制,它在动物生物学的许多方面都有作用。基于生物医学背景,几种信号启动了凋亡机制,包括前列腺素(PGs)。基于这些信息,我们提出了一个假设,即 PGs 同样会诱导昆虫细胞系发生凋亡。我们使用了三种 Spodoptera frugiperda 细胞系,包括从中枢神经系统中分离的两个新建立的 BCIRL-SfNS-0518B-YL 和从脂肪体中分离的 BCIRL-Sf4FB-0614-SGS,以及市售的 Sf9 细胞。使用细胞凋亡动力学试剂盒,我们发现用 15 或 20 μM PGA 处理 SfNS 细胞 18 小时会导致细胞数量减少,同时凋亡和死亡细胞数量增加。类似地,用 10 μM PGA(24 小时)处理会导致凋亡细胞数量显著增加,通过流式细胞仪上的末端脱氧核苷酸转移酶 dUTP 缺口末端标记测定法得到了证实。PGA 处理的影响随剂量增加而增加,因为我们在 PGA 处理后 24 小时记录到约 20%的凋亡,而在 30 μM 处理后 24 小时记录到约 34%的凋亡。PGA 处理在 12 小时时导致凋亡特异性半胱氨酸天冬氨酸蛋白酶-1、-2、-3 和 -5 的信使 RNA(mRNA)编码增加 10-30 倍,在 24 小时时导致 caspase-1 和 -2 的 mRNA 编码增加 40-60 倍,caspase-3 和 -5 的 mRNA 编码增加 10 倍。这些发现强烈支持我们的假设,即 PGs 诱导昆虫细胞系发生凋亡,并证实 PG 在昆虫生物学中的另一种作用。