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. 2013 Dec 11;14(12):24087-96.
doi: 10.3390/ijms141224087.

Radiation-sensitising effects of antennapedia proteins (ANTP)-SmacN7 on tumour cells

Li Qing Du, Yan Wang, Chang Xu, Jia Cao, Qin Wang, Hui Zhao, Fei Yue Fan, Bing Wang, Takanori Katsube, Sai Jun Fan, Qiang Liu  1
Affiliations

Radiation-sensitising effects of antennapedia proteins (ANTP)-SmacN7 on tumour cells

Li Qing Du et al. Int J Mol Sci. .

Abstract

The objective of this study was to investigate the underlying mechanisms behind the radiation-sensitising effects of the antennapedia proteins (ANTP)-smacN7 fusion protein on tumour cells. ANTP-SmacN7 fusion proteins were synthesised, and the ability of this fusion protein to penetrate cells was observed. Effects of radiation on the expression of X-linked inhibitor of apoptosis protein (XIAP) were detected by western blotting. The radiation-sensitising effects of ANTP-SmacN7 fusion proteins were observed by a clonogenic assay. The effects of drugs and radiation on tumour cell apoptosis were determined using Annexin V/FITC double staining. Changes in caspase-8, caspase-9 and caspase-3 were detected by western blot before and after ANTP-SmacN7 inhibition of XIAP. The ANTP-SmacN7 fusion protein could enter and accumulate in cells; in vitro XIAP expression of radiation-induced tumour cells was negatively correlated with tumour radiosensitivity. The ANTP-SmacN7 fusion protein promoted tumour cell apoptosis through the activation of caspase3. ANTP-SmacN7 fusion protein may reduce tumour cell radioresistance by inducing caspase3 activation.

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Figures

Figure 1.
Figure 1.
Relationship between XIAP expression of radiation-induced tumour cells and tumour radiosensitivity (a) Colony number of tumour cells after exposure to γ-ray; and (b) Western blot of the HeLa, H460, EC109 and 7901 tumour cell lines probed with antibodies to XIAP and β-actin.
Figure 2.
Figure 2.
Comparison of cell transduction capabilities of SmacN7 and the ANTP-SmacN7 fusion protein. Blue shows nuclear staining (DAPI), green (FITC) shows carboxyl terminus-labelled ANTP, SmacN7 and ANTP-SmacN7.
Figure 3.
Figure 3.
Radiation sensitising effects of ANTP-SmacN7.
Figure 4.
Figure 4.
Effects of ANTP-SmacN7 on radiation-induced apoptosis (a). The apoptotic index of EC109 cells after treatment with ANTP, SmacN7, and ANTP-SmacN7 combined with radiation; and (b) Graph of the flow cytometry results.
Figure 5.
Figure 5.
Effects of ANTP-SmacN7 on the tumour cell apoptotic pathway.
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