Expression And Breast Cancer Cell Inhibiting Effect Of Recombinant Camel Lactoferrin Peptides

Lactoferrin is a glycoprotein with broad range of antimicrobial activates. Recent studies indicatethat the anti-cancer effect of LF is attributed mostly to its ability to inhibit tumor cell proliferation,enhance apoptosis or necrosis in cancer cells. It is well documented that LF contains severalpeptides with multiple biological functions. In this study, we expressed a chimeric CamelLactoferrin peptide (cLF36) in Human Embryonic Kidney cells and tested its cytotoxic activity on ahuman breast cancer cell line Mcf7.The cLF36 sequence was codon optimized and synthesized.Vector harboring cLF36 sequence was digested, and ligated into the pcDNA3.1+ vector, thentransformed into TOP10 bacterium. The recombinant vector transfected into the HEK293 cells,using calcium phosphate method and were selected by G418 antibiotic. The culture medium uponcells was collected for SDS-PAGE analysis. MTT assay was used to examine the cytotoxicity of cLF36against of Mcf7 cells. Mcf7 cells were treated with medium containing the peptide and the culturemedium without the peptide and with DOXORUBICIN served as the negative and positive controlrespectively. The results of sequencing showed that cLF36 fragment was successfully cloned.Expression of the cLF36 in HEK293 cells was evaluated by SDS-PAGE, suggested that the protein wasproperly translated and secreted into medium culture. The result of MTT assay, showed significantincrease the rate of cell death when the cells treated with the medium containing cLF36 peptide.Treatment with cLF36 decreased viability level of Mcf7 cells to 32%. The main mechanism of anticanceractivity of lactoferrin peptides is still unknown, but it has been shown that lactoferrin cantarget the negatively charged of cancer cells. Our results revealed that cLF36 is cytotoxic for Mcf7cells, and it has considerable potential for therapeutic use in the treatment of breast cancer.