Shen Ying, De Schutter Kristof, Walski Tomasz, Van Damme Els J M, Smagghe Guy
Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, 9000, Ghent, Belgium.
Laboratory of Biochemistry and Glycobiology, Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, 9000, Ghent, Belgium.
In Vitro Cell Dev Biol Anim. 2017 Sep;53(8):691-698. doi: 10.1007/s11626-017-0176-8. Epub 2017 Jul 11.
The fungal lectin purified from Sclerotinia sclerotiorum, further referred to as Sclerotinia sclerotiorum agglutinin or SSA, possesses insecticidal activity against important pest insects such as pea aphids (Acyrthosiphon pisum). This paper aims at a better understanding of its activity at cellular level. Therefore, different insect cell lines were treated with SSA. These cell lines were derived from different tissues and represent the three major orders of insects important in agriculture: CF-203 (midgut Choristoneura fumiferana, Lepidoptera), GUTAW1 (midgut, Helicoverpa zea, Lepidoptera), High5 cells (ovary, Trichoplusia ni, Lepidoptera), Sf9 (ovary cells from Spodoptera frugiperda, Lepidoptera), S2 (hemocyte, Drosophila melanogaster, Diptera), and TcA (whole body, Tribolium castaneum, Coleoptera). Although the sensitivity to SSA differs between the cell lines, SSA clearly showed toxicity in all six cell lines with median effect concentrations (EC) ranging between 9 and 42 μg/ml. An in-depth analysis of the mechanism of uptake in the cells revealed superior amounts of FITC-SSA at the membrane of CF-203 cells compared to Sf9 cells, while a similar small amount of SSA was internalized in both cell lines. Pre-incubation with the clathrin-mediated endocytosis inhibitor phenylarsine oxide inhibited the internalization of SSA into the CF-203 and Sf9 cells with a respective reduction of 6- and 1.7-fold. The data are discussed in relation to the importance of cellular uptake mechanism for SSA binding and cytotoxicity.
从核盘菌中纯化得到的真菌凝集素，以下简称核盘菌凝集素或SSA，对豌豆蚜（Acyrthosiphon pisum）等重要害虫具有杀虫活性。本文旨在更好地了解其在细胞水平上的活性。因此，用SSA处理了不同的昆虫细胞系。这些细胞系来源于不同组织，代表了农业中重要的三个主要昆虫目：CF-203（云杉卷叶蛾中肠细胞，鳞翅目）、GUTAW1（玉米螟中肠细胞，鳞翅目）、High5细胞（粉纹夜蛾卵巢细胞，鳞翅目）、Sf9（草地贪夜蛾卵巢细胞，鳞翅目）、S2（黑腹果蝇血细胞，双翅目）和TcA（赤拟谷盗全身细胞，鞘翅目）。尽管各细胞系对SSA的敏感性不同，但SSA在所有六个细胞系中均表现出毒性，中位效应浓度（EC）在9至42μg/ml之间。对细胞摄取机制的深入分析表明，与Sf9细胞相比，CF-203细胞膜上的FITC-SSA含量更高，而两个细胞系内化的SSA量相似。用网格蛋白介导的内吞作用抑制剂氧化苯胂预孵育可抑制SSA进入CF-203和Sf9细胞，内化量分别降低6倍和1.7倍。本文结合细胞摄取机制对SSA结合和细胞毒性的重要性对数据进行了讨论。
