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.
Phleomycin是一种糖肽类抗生素，是一种DNA链断裂（DNA-SB）诱导剂，可用于筛选基因工程细胞[1]。Phleomycin是在链霉菌中发现的，与博来霉素 （Bleomycin）类似，它能够结合并插入DNA，破坏双螺旋结构的完整性[2]。Phleomycin对抗性链球菌菌株具有诱变作用[3]。
在体外，Phleomycin（100-400μg/mL）处理HeLa细胞0-24h，以时间和剂量依赖性方式抑制了细胞增殖，诱导了细胞毒性，诱导了细胞色素c的初始释放、Bcl-XL、ENDOG、DAXX和MDM2的下调以及CASP和BID的上调[4]。Phleomycin（0.2μg/mL）处理微拟球藻（Nannochloropsis oceanica）细胞48h，引起了藻细胞DNA的随机双链断裂，提高了活性潮霉素B抗性基因（aph7）的转化效率[5]。Phleomycin（0-200ng/mL）处理来自不同突变体的鼠胚胎成纤维细胞（MEFs）48h，显著降低了缺乏XRCC4、DNA-Ligase IV或Xlf的MEFs的存活率，但对来自PAXX突变体的MEFs和野生型MEFs无显著影响[6]。