Bo Tang1, Da-Wei He, Dian Li, Wen-Hao Guo, Dan Zhang, Guang-Hui Wei. 1. Department of Urinary Surgery, Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing 400014, China. E-mail: ljsmilekiss@126.com.
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 apoptosis-related 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.
OBJECTIVE: To investigate the effect of outer membrane vesicles derived from Escherichia coli on proliferation, apoptosis and migration of humanneuroblastoma 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. Humanneuroblastoma 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 apoptosis-related 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.
Authors: Martina Bielaszewska; Christian Rüter; Lisa Kunsmann; Lilo Greune; Andreas Bauwens; Wenlan Zhang; Thorsten Kuczius; Kwang Sik Kim; Alexander Mellmann; M Alexander Schmidt; Helge Karch Journal: PLoS Pathog Date: 2013-12-12 Impact factor: 6.823