%0 Journal Article %A Liu, Xiufeng %A Liu, Xintong %A Sunchen, Suwen %A Liu, Meixia %A Shen, Chen %A Wu, Juanjuan %A Zhao, Wanli %A Yu, Boyang %A Liu, Jihua %D 2017 %T A novel tumor-activated ALA fusion protein for specific inhibition on the growth and invasion of breast cancer cells MDA-MB-231 %U https://tandf.figshare.com/articles/journal_contribution/A_novel_tumor-activated_ALA_fusion_protein_for_specific_inhibition_on_the_growth_and_invasion_of_breast_cancer_cells_MDA-MB-231/5632279 %R 10.6084/m9.figshare.5632279 %2 https://ndownloader.figshare.com/files/9809026 %K Fusion protein %K tumor activate %K uPA %K AGAP %K apoptosis %X

Objective: The aim of this research was to develop a novel ALA fusion protein for target to the malignant cells surface with high uPAR expression and locally release of the scorpion toxin AGAP in an uPA-cleavable manner. It will provide an effective approach for controlled release of the peptide toxins to treat cancerous cells.

Methods: The ALA fusion proteins were expressed in pichia pastoris, and the recombinant proteins were purified by Ni-NTA affinity chromatography. The proteins were added to human breast cancer cells (MDA-MB-231) and human embryonic kidney cells (HEK-293) in order to investigate the characteristic of selective targeting and releasing of scorpion toxin AGAP in cancer cells with high uPAR expression. The inhibitory effect of ALA on MDA-MB-231, MCF7, LO2 and HEK-293 was evaluated by MTT assay. Moreover, the antiproliferation mechanism of ALA was determined by flow cytometric and western blot analysis.

Results: The results showed that ALA could target MDA-MB-231 cells and the scorpion toxin AGAP could be released with high efficiency and selectivity. ALA inhibited the growth and invasion of breast cancer cells MDA-MB231. Also, cell apoptosis pathway was found to be associated with the inhibition mechanism of ALA according to the data of flow cytometric and western blot analysis. Therefore, ALA could be a novel antitumor candidate for targeting treatment of malignant cell.

Conclusions: This study successfully demonstrated that fusion of biotoxins with tumor target domain could provide a simple yet effective way to delivery of peptide or protein drugs.

%I Taylor & Francis