Antiproliferative effect of azadirachtin A on Spodoptera litura Sl-1 cell line through cell cycle arrest and apoptosis induced by up-regulation of p53
Antiproliferative effect of azadirachtin A on Spodoptera litura Sl-1 cell line through cell cycle arrest and apoptosis induced by up-regulation of p53
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印楝素 A 通过 p53 上调诱导的细胞周期阻滞和细胞凋亡对斜纹夜蛾 Sl-1 细胞系的抗增殖作用
DOI：
10.1016/j.pestbp.2010.08.002
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发表时间：
2011-01-01
影响因子：
4.7
通讯作者：
Zhong, Guo-Hua
中科院分区：
文献类型：
--
作者：
Huang, Jing-Fei;Shui, Ke-Juan;Zhong, Guo-Hua
为了阐明植物源杀虫剂诱导细胞周期停滞和凋亡的机制，本研究以斜纹夜蛾Sl-1细胞为材料，研究了印楝素A对细胞增殖的抑制和诱导细胞凋亡的作用。采用四甲基偶氮唑盐比色法、流式细胞仪、荧光显微镜观察、免疫印迹等方法检测细胞增殖、细胞周期分裂、核有丝分裂和凋亡形态及相关蛋白的表达。印防素A以0.1mU g/mL、0.5mU g/m L和1.0mU g/m L作用于SL-1细胞48h后，细胞生长呈剂量依赖性抑制，抑制率分别为18.95%、33.45%和41.00%。SL-1细胞周期停滞于G2/M期，Cyclin B1表达降低。0.1、0.5、1.0mU g/m L印木素A作用48h后，细胞凋亡率分别为19.71%、23.40%和24.91%，细胞凋亡率随caspase-3活性的升高而升高。在细胞凋亡和细胞周期停滞发生之前，就观察到了P53的上调。这些结果表明，印楝素A(C)2010 Elsevier Inc.处理昆虫培养细胞时，p53蛋白参与了细胞周期停滞、诱导凋亡和抑制细胞增殖。版权所有。
In order to illustrate the mechanism of cell cycle arrest and apoptosis induced by botanical insecticide, the effects of cell proliferation inhibition and apoptosis induction by azadirachtin A were investigated in Spodoptera litura cell line Sl-1 in vitro. The cell proliferation, cell cycle division, mitotic and apoptotic figure of nucleus, and the expression of related proteins were detected by a variety of methods, including MTT assay, flow cytometry, fluorescence microscope observation, and western blot technique. Sl-1 cells grew in a dose-dependent inhibition manner and the proliferation inhibition rates were 18.95%, 33.45% and 41.00%, after 48 h of treatment with azadirachtin A at the concentrations of 0.1 mu g/mL, 0.5 mu g/mL and 1.0 mu g/mL, respectively. Sl-1 cell cycle was arrested in G2/M phase with lower expression of cyclin B1. The apoptosis rates were found to be 19.71%, 23.40% and 24.91% after 48 h of treatment with 0.1, 0.5, and 1.0 mu g/mL of azadirachtin A. which increased with the activities of caspase-3. The up-regulation of p53 was observed before apoptosis and cell cycle arrest occurred. These findings suggest that p53 protein was involved in cell cycle arrest, apoptosis induction and cell proliferation inhibition when the insect cultured cells were treated with azadirachtin A. (C) 2010 Elsevier Inc. All rights reserved.