Phleomycin is a glycopeptide antibiotic and a DNA strand break (DNA-SB) inducer that can be used to select genetically-engineered cells[1]. Phleomycin is a molecule found in Streptomyces, and similar to bleomycin, can bind to and insert into DNA, disrupting the integrity of the double helix structure[2]. Phleomycin has a mutagenic effect on resistant Streptococcus strains[3].
In vitro, treatment of HeLa cells with Phleomycin (100-400μg/mL) for 0-24h inhibited cell proliferation in a time- and dose-dependent manner, induced cytotoxicity, induced the initial release of cytochrome c, downregulation of Bcl-XL, ENDOG, DAXX, and MDM2, and upregulation of CASP and BID[4]. Treatment of Nannochloropsis oceanica cells with Phleomycin (0.2μg/mL) for 48h caused random double-strand breaks in the DNA of the algal cells and increased the transformation efficiency of the active hygromycin B resistance gene (aph7)[5]. Phleomycin (0-200ng/mL) treatment of mouse embryonic fibroblasts (MEFs) from different mutants for 48h significantly reduced the survival rate of MEFs lacking XRCC4, DNA-Ligase IV or Xlf, but had no significant effect on MEFs from PAXX mutants and wild-type MEFs[6].
References:
[1] Ma X, Pan K, Zhang L, et al. Genetic transformation of Nannochloropsis oculata with a bacterial phleomycin resistance gene as dominant selective marker[J]. Journal of Ocean University of China, 2016, 15: 351-356.
[2] Stokowa-Sołtys K, Dzyhovskyi V, Wieczorek R, et al. Phleomycin complex–Coordination mode and in vitro cleavage of DNA[J]. Journal of inorganic biochemistry, 2019, 195: 71-82.
[3] Van Peer A F, De Bekker C, Vinck A, et al. Phleomycin increases transformation efficiency and promotes single integrations in Schizophyllum commune[J]. Applied and Environmental Microbiology, 2009, 75(5): 1243-1247.
[4] Hwang J, Kim Y Y, Huh S, et al. The Time‐Dependent Serial Gene Response to Zeocin Treatment Involves Caspase‐Dependent Apoptosis in HeLa Cells[J]. Microbiology and immunology, 2005, 49(4): 331-342.
[5] Zhang Z, Guo L, Liu H, et al. Zeocin treatment significantly elevated transformation efficiency of Nannochloropsis oceanica[J]. Journal of Applied Phycology, 2022, 34(3): 1587-1594.
[6] Abramowski V, Etienne O, Elsaid R, et al. PAXX and Xlf interplay revealed by impaired CNS development and immunodeficiency of double KO mice[J]. Cell Death & Differentiation, 2018, 25(2): 444-452.