Induction of Apoptosis on K562 Cell Line and Double Strand Breaks on Colon Cancer Cell Line Expressing High Affinity Receptor for Granulocyte Macrophage-Colony Stimulating factor (GM-CSF)

Authors
Mehryar Habibi Roudkenar
Saeid Bouzari
Yoshikazu Kuwahara
Amaneh Mohammadi
Taisuke Baba
Mana Oloomi
Manabu Fukumoto
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Abstract
Background: Immunotoxins are comprised of both the cell targeting and the cell killing moieties. We previously established a new immunotoxin, i.e. Shiga toxin granulocyte macrophage-colony stimulating factor (StxA1-GM-CSF), comprises of catalytic domain of Stx, as a killing moiety and GM-CSF, as a cell targeting moiety. In this study, the ability of the immunotoxin to induce apoptosis and double strand breaks (DSB) on different cell lines was investigated. Methods: The recombinant hybrid protein was expressed in bacterial expression system and purified with nickel-nitrilotriacetate acid resin. The K562 (erythroid leukemia) cell line and LS174 (colon carcinoma) were used in this study. The neutral comet assay was carried out for the detection of DSB and Hoechst staining was performed for apoptosis. Results: StxA1-GM-CSF effectively induced apoptosis on K562 cell line and DNA Double Strand Break (DSB) were observed on colon cancer cell line treated with StxA1-GM-CSF. Conclusion: This novel action i.e. DNA damage might be a relevant mechanism of action for StxA1-GM-CSF that is designed to act as immunotoxin, although further investigation is required.
Keywords
Shiga toxin granulocyte macrophage-colony stimulating factor (StxA1-GM-CSF)
Cancer cells
Apoptosis
Double strand