Study on induction of apoptosis on HeLa and Vero cells by recombinant shiga toxin and its subunits.
Verotoxin (VT) or shiga toxin (Stx) produced by enterohemorrhagic Escherichia coli (EHEC) and Shigella dysenteriae is AB5 holotoxin with potent protein synthesis inhibitor. VT can induce both apoptosis and necrosis depending on the cell type, it has been shown that VT-induced apoptosis and cytotoxicity are distinct processes, and the A subunit can be necessary for apoptosis. In other words, the precise role of each subunit in apoptosis signaling has yet to be established. In this study, induction of apoptosis has been examined by using both recombinant A and B subunits, and recombinant Stx (rStx) with different doses in HeLa and Vero cells. For this purpose, the polymyxin B extract of constructs expressing A, B and AB5 recombinant proteins was used. Therefore, amounts greater than normally reported were used to induce desire effects on cell lines. The apoptotic effect of A and B subunits appear at higher doses than that of rStx. The highest apoptotic effect was observed for rStx at low concentration, compared to A and B subunits. A or B subunits separately cannot induce the signaling pathway stimulated by holotoxin though A subunit, does induce laddering pattern similar to holotoxin. We concluded that both subunits are important in complete death signaling pathway. Since different concentration of A and B subunits and rStx was required in different assay, therefore, it could be emphasized that cell death or even apoptosis caused by either of the subunits or holotoxin depends on sensitivity or specificity of the assay and cell types used.
Bouzari
Saeid
S
Molecular Biology Unit, Pasteur Institute of Iran, Pasteur Ave., 13164, Tehran, Iran.
Oloomi
Mana
M
Molecular Biology Unit, Pasteur Institute of Iran, Pasteur Ave., 13164, Tehran, Iran. manaoloomi@yahoo.com.
Azadmanesh
Kayhan
K
Molecular Biology Unit, Pasteur Institute of Iran, Pasteur Ave., 13164, Tehran, Iran.