ABSTRACT
Background: Arsenic trioxide [ATO] has been reported as an effective anti-cancer and a US Food and Drug Administration [FDA] approved drug for treatment of some cancers. The aim of this study was to determine the underlying apoptosis molecular and cellular mechanisms of ATO in the presence or absence of ionizing radiation [IR] in vitro in the glioblastoma multiforme [GBM] cell line, U87MG
Methods: Cells were treated by different concentrations of ATO either in presence or absence of IR. Viability and apoptosis pathway of both treated and control groups were evaluated using MTT assay and the expression analysis of Box, Bcl-2, and caspase-3 genes, respectively. All treatments were performed on 100-ujm diameter spheroids
Results: Results showed a significant reduction in the survival of the cells in all treated groups. As expected, cell survival was much less in combination treatment than treatment with only ATO. Moreover, combination therapy made Box and caspase-3 up-regulated and Bcl-2 down-regulated
Conclusion: ATO and radiation had a synergistic apoptotic effect on GBM cells by up-regulation of caspase-3 and alteration of the Bax-Bcl-2 balance; therefore, ATO may act as a potential anti-cancer agent against GBM cells through triggering the mitochondria! pathway of apoptosis
Subject(s)
Journal Article , Apoptosis/radiation effects , Arsenicals/therapeutic use , Oxides/therapeutic use , Radiation, Ionizing , In Vitro Techniques , Glioblastoma , Cell Line, TumorABSTRACT
Background: As a drug target and an antigenic agent, HIV-1 protease [HIV-1 PR] is at the center of attention for designing anti-AIDS inhibitors and diagnostic tests. In previous studies, the production of the recombinant protease has been faced with several difficulties; therefore, the aims of this study were the easy production, purification of the soluble form of protease in E. coli and investigation of its immunoreactivity
Methods: Protease coding region was isolated from the serum of an infected individual, amplified by RT-PCR and cloned into PTZ57R using TA-cloning. Protease coding frame was isolated by PCR and cloned in pET102/D. TOPO expression vector and cloned protease was expressed in Escherichia coli [E. coli] BL21. Produced recombinant protein was purified by affinity Ni-NTA column and protein concentration was checked by BCA protein assay kit. Subsequently, immunoreactivity of recombinant protease [rPR] was assayed by Western blotting and ELISA
Results: Cloning of the HIV protease by TOPO cloning system in pET102/D.TOPO was confirmed with PCR and sequencing. The concentration range of purified recombinant protein was 85 to 100 micro g/ml. Immunogenicity of rPR was confirmed by Western blotting and ELISA
Conclusion: Soluble production of recombinant HIV-1 protease [HIV-1 rPR] was performed successfully. This recombinant protein disclosed 86% specificity and 90% sensitivity in immunoassay tests