[1]唐波,何大维,李典,等.大肠杆菌胞外囊泡对人神经母细胞瘤SK-N-SH细胞增殖、凋亡和迁移的影响[J].南方医科大学学报,2018,(03):334.
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大肠杆菌胞外囊泡对人神经母细胞瘤SK-N-SH细胞增殖、凋亡和迁移的影响()
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《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2018年03期
页码:
334
栏目:
出版日期:
2018-04-03

文章信息/Info

Title:
Effect of outer membrane vesicles derived from Escherichia coli on proliferation, apoptosis and migration of human neuroblastoma SK-N-SH cells in vitro
作者:
唐波何大维李典郭文浩张丹魏光辉
关键词:
SK-N-SH细胞大肠杆菌胞外囊泡增殖凋亡迁移
Keywords:
neuroblastoma SK-N-SH cells outer membrane vesicles Escherichia coli cell proliferation apoptosis migration
摘要:
目的研究大肠杆菌胞外囊泡(E.coli-OMVs)对人神经母细胞瘤SK-N-SH细胞增殖、凋亡和迁移的影响。方法采用差速 离心法提取大肠杆菌胞外囊泡,观察囊泡形态并测定囊泡粒径范围。随机分为4组:正常对照组、胞外囊泡低剂量组(100 μg/mL E.coli-OMVs)、胞外囊泡中剂量组(500 μg/mL E.coli-OMVs)和胞外囊泡高剂量组(1000 μg/mL E.coli-OMVs)。MTT检测各组 细胞增殖能力;TUNEL和Western blot分别检测各组细胞凋亡情况及凋亡相关蛋白caspase3的表达;Transwell检测各组细胞迁 移能力。结果透射电镜下E.coli-OMVs呈双层膜、中空、圆形或椭圆形,其内含低密度或高密度物质,具有明显的异质性,粒径 分析结果表明其直径分布在30~450 nm。与对照组相比,经E.coli-OMVs干预后,SK-N-SH细胞增殖能力显著降低;细胞凋亡 增加,caspase3蛋白表达水平明显上调;而迁移能力显著降低(P<0.05)。结论E.coli-OMVs对SK-N-SH细胞具有直接杀伤作 用,有望成为难治性神经母细胞瘤的治疗药物。
Abstract:
Objective To investigate the effect of outer membrane vesicles derived from Escherichia coli on proliferation, apoptosis and migration of human neuroblastoma SK-N-SH cells in vitro. Methods The outer membrane vesicles (OMVs) were obtained from wild-type Escherichia coli with ultracentrifugation method, and the morphology of the OMVs was observed by transmission electron microscopy and the vesicle diameter was determined using MALVERN ZEN3690. Human neuroblastoma SK-N-SH cells were treated with the OMVs at low (100 μg/mL), moderate (500 μg/mL) and high (1000 μg/mL) doses, and 24, 48 and 72 h later, the cell proliferation activity was detected with MTT assay. The expressions of apoptosisrelated marker caspase-3 was detected using Western blotting, and TUNEL assay was performed to detect the cell apoptosis. The migration capacity of SK-N-SH cells was evaluated using Transwell migration assay. Results The isolated OMVs showed a circular or elliptical hollow structure with double-layer membrane and a diameter range of 30-450 nm. Compared with the control cells, SK-N-SH cells treated with the OMVs showed significantly lowered cell proliferation capacity with enhanced expression of caspase-3. Treatment of the cells with the OMVs resulted in increased cell apoptosis and significantly lowered migration capacity (P<0.05). Conclusion The OMVs derived from Escherichia coli can produce cytotoxicity against SK-N-SH cells and might serve as a therapeutic agent for refractory neuroblastoma.
更新日期/Last Update: 1900-01-01