The use of fluorescent halo assay and acridine orange flow cytometry to predict intrinsic radiosensitivity, radiation-induced dna damage and repair in human tumor cells

Author(s):  
Karl King ◽  
Ryuji Higashikubo ◽  
Joseph Roti Roti
Viruses ◽  
2018 ◽  
Vol 10 (8) ◽  
pp. 393 ◽  
Author(s):  
Liqian Zhu ◽  
Xiaotian Fu ◽  
Chen Yuan ◽  
Xinyi Jiang ◽  
Gaiping Zhang

Bovine herpesvirus 1 (BoHV-1) is an important pathogen of cattle that causes lesions in mucosal surfaces, genital tracts and nervous systems. As a novel oncolytic virus, BoHV-1 infects and kills numerous human tumor cells. However, the mechanisms underlying the virus-induced cell damages are not fully understood. In this study, we demonstrated that virus infection of MDBK cells induced high levels of DNA damage, because the percentage of comet tail DNA (tailDNA%) determined by comet assay, a direct indicator of DNA damage, and the levels of 8-hydroxyguanine (8-oxoG) production, an oxidative DNA damage marker, consistently increased following the virus infection. The expression of 8-oxoguanine DNA glycosylase (OGG-1), an enzyme responsible for the excision of 8-oxoG, was significantly decreased due to the virus infection, which corroborated with the finding that BoHV-1 infection stimulated 8-oxoG production. Furthermore, the virus replication in human tumor cells such as in A549 cells and U2OS cells also induced DNA damage. Chemical inhibition of reactive oxidative species (ROS) production by either ROS scavenger N-Acetyl-l-cysteine or NOX inhibitor diphenylene iodonium (DPI) significantly decreased the levels of tailDNA%, suggesting the involvement of ROS in the virus induced DNA lesions. Collectively, these results indicated that BoHV-1 infection of these cells elicits oxidative DNA damages, providing a perspective in understanding the mechanisms by which the virus induces cell death in both native host cells and human tumor cells.


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George Vartholomatos ◽  
Nikolaos Monokrousos ◽  
...  

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