Bacterial delivery of the anti-tumor azurin-like protein Laz to glioblastoma cells
Abstract Salmonella typhimurium VNP-20009 (VNP) is a non-pathogenic attenuated strain, which, as a facultative anaerobe, preferentially accumulates in hypoxic regions of solid tumors. Here, VNP was utilized as a delivery vehicle of the anti-tumor protein Lipidated azurin, Laz, which is produced by t...
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doaj-fc6aebb766394a5db9fcb2afe01c22432020-11-25T01:53:44ZengSpringerOpenAMB Express2191-08552020-03-0110111010.1186/s13568-020-00995-8Bacterial delivery of the anti-tumor azurin-like protein Laz to glioblastoma cellsManar Mansour0Shehab Ismail1Khaled Abou-Aisha2Department of Microbiology and Immunology, The German University in Cairo (GUC)The Cancer Research Institute CRUK Beatson InstituteDepartment of Microbiology and Immunology, The German University in Cairo (GUC)Abstract Salmonella typhimurium VNP-20009 (VNP) is a non-pathogenic attenuated strain, which, as a facultative anaerobe, preferentially accumulates in hypoxic regions of solid tumors. Here, VNP was utilized as a delivery vehicle of the anti-tumor protein Lipidated azurin, Laz, which is produced by the meningitis-causing bacterium Neisseria meningitides. In brain cancer cells, Laz has been demonstrated to induce apoptosis through an interaction with the tumor suppressor protein p53. In this study, the laz gene, including its signal sequence, was cloned downstream of a hypoxia inducible promoter (HIP-1), before being electroporated into VNP. Successful ectopic expression and export of the Laz protein by VNP under hypoxic conditions were confirmed by Western blot analysis of the cell-free culture medium. Effective expression of Laz by VNP was investigated in two glioblastoma cell lines: LN-229 and U-373, with the latter line carrying a mutated version of p53; as well as in the breast cancer line MCF-7. Cytotoxicity of the VNP-Laz was assessed by determining the fluorescence of the apoptotic marker caspases 3/7. Compared to the purified Laz, VNP-Laz, significantly induced apoptosis in MCF-7, LN-229 and, to a much lower extent in U-373 cells, suggesting a p53-linked mechanism. Our results might represent a new approach of targeted gene delivery and suggest a potential application in brain tumor therapy.http://link.springer.com/article/10.1186/s13568-020-00995-8HypoxiaSalmonella VNP20009Azurin-like proteinGlioblastoma |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Manar Mansour Shehab Ismail Khaled Abou-Aisha |
spellingShingle |
Manar Mansour Shehab Ismail Khaled Abou-Aisha Bacterial delivery of the anti-tumor azurin-like protein Laz to glioblastoma cells AMB Express Hypoxia Salmonella VNP20009 Azurin-like protein Glioblastoma |
author_facet |
Manar Mansour Shehab Ismail Khaled Abou-Aisha |
author_sort |
Manar Mansour |
title |
Bacterial delivery of the anti-tumor azurin-like protein Laz to glioblastoma cells |
title_short |
Bacterial delivery of the anti-tumor azurin-like protein Laz to glioblastoma cells |
title_full |
Bacterial delivery of the anti-tumor azurin-like protein Laz to glioblastoma cells |
title_fullStr |
Bacterial delivery of the anti-tumor azurin-like protein Laz to glioblastoma cells |
title_full_unstemmed |
Bacterial delivery of the anti-tumor azurin-like protein Laz to glioblastoma cells |
title_sort |
bacterial delivery of the anti-tumor azurin-like protein laz to glioblastoma cells |
publisher |
SpringerOpen |
series |
AMB Express |
issn |
2191-0855 |
publishDate |
2020-03-01 |
description |
Abstract Salmonella typhimurium VNP-20009 (VNP) is a non-pathogenic attenuated strain, which, as a facultative anaerobe, preferentially accumulates in hypoxic regions of solid tumors. Here, VNP was utilized as a delivery vehicle of the anti-tumor protein Lipidated azurin, Laz, which is produced by the meningitis-causing bacterium Neisseria meningitides. In brain cancer cells, Laz has been demonstrated to induce apoptosis through an interaction with the tumor suppressor protein p53. In this study, the laz gene, including its signal sequence, was cloned downstream of a hypoxia inducible promoter (HIP-1), before being electroporated into VNP. Successful ectopic expression and export of the Laz protein by VNP under hypoxic conditions were confirmed by Western blot analysis of the cell-free culture medium. Effective expression of Laz by VNP was investigated in two glioblastoma cell lines: LN-229 and U-373, with the latter line carrying a mutated version of p53; as well as in the breast cancer line MCF-7. Cytotoxicity of the VNP-Laz was assessed by determining the fluorescence of the apoptotic marker caspases 3/7. Compared to the purified Laz, VNP-Laz, significantly induced apoptosis in MCF-7, LN-229 and, to a much lower extent in U-373 cells, suggesting a p53-linked mechanism. Our results might represent a new approach of targeted gene delivery and suggest a potential application in brain tumor therapy. |
topic |
Hypoxia Salmonella VNP20009 Azurin-like protein Glioblastoma |
url |
http://link.springer.com/article/10.1186/s13568-020-00995-8 |
work_keys_str_mv |
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