Anti-cancer Effect of Recombinant PI-Laterosporulin10 as a Novel Bacteriocin with Selective Cytotoxicity on Triple Negative Breast Cancer Cells

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, puri cation, and cytotoxic effects of recombinant PI-laterosporulin10 (LS10) a defensing-like peptide. PI is a transmembrane peptide that helps to increase penetration and speci city 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 E. 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 hours post-induction. After enzymatic cleavage, the purity of PI-LS10 was >300 µg/ ml as the nal 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 e ciency in targeting triple negative cells. Our results suggest that PI-LS10 had a signi cant 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 hours 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 con rmed.