Abstract
Anti-microbial peptides (AMPs) are important factors of the innate immune system which perform a comprehensive spectrum of biological activities. The present study aims to examine the expression, purification, and cytotoxic effects of recombinant PI–laterosporulin10 (LS10) a defensing-like peptide. PI is a transmembrane peptide that helps to increase penetration and specificity of peptide targeting. PI–LS10 sequence was optimized and fused to a Xa protease site to achieve higher levels of expression. Then Xa–PI–LS10 was cloned into the pET-32a (+) vector and expressed in Escherichia coli BL21 (DE3). The fusion protein with a molecular weight of about 25.3 kDa was analyzed using the SDS-PAGE. The maximum expression was observed 8 h post-induction. After enzymatic cleavage, the purity of PI–LS10 was > 300 µg/ ml as the final yield. The recombinant PI–LS10 showed anti-cancer characteristics against breast cancer cell lines. MCF7 (non-triple negative) and MDA-MB-231 (triple negative) cells were treated with different concentrations of the PI–LS10 in order to evaluate their cytotoxic activity and efficiency in targeting triple negative cells. Our results suggest that PI–LS10 had a significant cytotoxic effect against breast cancer cells in comparison to the normal cell line. Triple negative breast cancer cell (MDA-MB-231) proliferation inhibited higher in 72 h after treatment compare MCF7 cells. In conclusion, the PI–LS10 peptide was produced successfully and its favorable anti-cancer activity against a breast cancer cell line was confirmed.
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Acknowledgements
This research was financially supported by Shahrekord University (Grant No. 99GRN31M32855). We thank all cooperating with us in conduction of this project, particularly the respectful populates and officials of Shahrekord University.
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BR: performed in vitro experimental prepared initial draft and data collection. SR: coordinated the study, supervised the data collection and proof read manuscript, perform data analysis and article editing. BS and SF: prepared information on complementary literature and performed data analysis. FT: performs article editing and share in prepared initial draft.
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Rafizadeh, B., Reiisi, S., Saffar, B. et al. Anti-cancer Effect of Recombinant PI-Laterosporulin10 as a Novel Bacteriocin with Selective Cytotoxicity on Triple Negative Breast Cancer Cells. Int J Pept Res Ther 28, 143 (2022). https://doi.org/10.1007/s10989-022-10453-7
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DOI: https://doi.org/10.1007/s10989-022-10453-7