A-NGR fusion protein induces apoptosis in human cancer cells
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Date
2018-06-25
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Abstract
The NGR peptide is one of the well-known peptides for targeting tumor cells. It has the ability to target aminopeptidase
N (CD13) on tumor cells or the tumor vascular endothelium. In this study, the NGR peptide was used for
targeting A subunit of the Shiga toxin to cancer cells. The cytotoxic effect of the A-NGR fusion protein was
assessed on HT1080, U937, HT29 cancer cells and MRC-5 normal cells. For this purpose, cells were treated with
different concentrations of A-NGR (0.5-40 μg/ml). The evaluation of cell viability was achieved by MTT assay.
Apoptosis was determined by annexin-V/PI double staining flow cytometry. Alterations in the mRNA expression
of apoptosis - related genes were assessed by real time RT- PCR. The results showed that A-NGR fusion protein
effectively inhibited the growth of HT1080 and U937 cancer cells in comparison to negative control (PBS) but for
CD13-negative HT-29 cancer cells, only at high concentrations of fusion protein was inhibited growth recorded.
On the other hand, A-NGR had little cytotoxic effect on MRC-5 normal cells. The flow cytometry results showed
that A-NGR induces apoptosis. Furthermore, the results of real time RT-PCR revealed that A-NGR significantly
increases the mRNA expression of caspase 3 and caspase 9. Conclusively, A-NGR fusion protein has the ability
of targeting CD13-positive cancer cells, the cytotoxic effect on CD13-positive cancer cells as well as has low
cytotoxic effect on normal cells.
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Keywords
Shiga toxin, NGR peptide, Apoptosis, Cytotoxicity