Assessment of antimicrobial (Host Defense) peptides as anti-cancer agents

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Journal titleMethods in Molecular Biology
Pages159170; # of pages: 12
Subjectantimicrobial cationic peptide; antineoplastic agent; chromium 51; lactate dehydrogenase; single stranded DNA; thymidine; adherent cell; apoptosis; cancer cell; cell death; cell suspension; cell transformation; clonogenic assay; cytotoxicity; DNA content; flow cytometry; host resistance; necrosis
AbstractCationic antimicrobial (host defense) peptides (CAPs) are able to kill microorganisms and cancer cells, leading to their consideration as novel candidate therapeutic agents in human medicine. CAPs can physically associate with anionic membrane structures, such as those found on cancer cells, causing pore formation, intracellular disturbances, and leakage of cell contents. In contrast, normal cells are less negatively-charged and are typically not susceptible to CAP-mediated cell death. Because the interaction of CAPs with cells is based on charge properties rather than cell proliferation, both rapidly dividing and quiescent cancer cells, as well as multidrug-resistant cancer cells, are targeted by CAPs, making CAPS potentially valuable as anti-cancer agents. CAPs often exist as families of peptides with slightly different amino acid sequences. In addition, libraries of synthetic peptide variants based on naturally occurring CAP templates can be generated in order to improve upon their action. High-throughput screens are needed to quickly and efficiently assess the suitability of each CAP variant. Here we present the methods for assessing CAP-mediated cytotoxicity against cancer cells (suspension and adherent) and untransformed cells (measured using the tritiated thymidine-release or MTT assay), and for discriminating between cell death caused by necrosis (measured using lactate dehydrogenase-or 51Cr- release assays), or apoptosis and necrosis (single-stranded DNA content measured by flow cytometry). In addition the clonogenic assay, which assesses the ability of single transformed cells to multiply and produce colonies, is described. © 2014 Springer Science+Business Media, LLC.
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AffiliationNational Research Council Canada (NRC-CNRC); NRC Institute for Marine Biosciences (IMB-IBM)
Peer reviewedYes
NPARC number21270839
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Record identifier7b0660d6-2cdc-4291-87f0-63d1d0a256ce
Record created2014-02-17
Record modified2016-05-09
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