Adriamycin induces H2AX phosphorylation in human spermatozoa

Asian J Androl. 2008 Sep;10(5):749-57. doi: 10.1111/j.1745-7262.2008.00400.x.

Abstract

Aim: To investigate whether adriamycin induces DNA damage and the formation of gammaH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa.

Methods: Human spermatozoa were treated with adriamycin at different concentrations. gammaH2AX was analyzed by immunofluorescent staining and flow cytometry and double-strand breaks (DSB) were detected by the comet assay.

Results: The neutral comet assay revealed that the treatment with adriamycin at 2 microg/mL for different times (0.5, 2, 8 and 24 h), or for 8 h at different concentrations (0.4, 2 and 10 microg/mL), induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of gH2AX was increased in a dose-dependent and time-dependant manner after the treatment of adriamycin. Adriamycin also induced the concurrent appearance of DNA maintenance/repair proteins RAD50 and 53BP1 with gammaH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3-kinase (PI3K) family, abolished the co-appearance of these two proteins with gammaH2AX.

Conclusion: Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/repair proteins as somatic cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acid Anhydride Hydrolases
  • Androstadienes / pharmacology
  • Antibiotics, Antineoplastic / pharmacology*
  • Cells, Cultured
  • Comet Assay
  • DNA Breaks, Double-Stranded / drug effects
  • DNA Damage
  • DNA Repair Enzymes / metabolism
  • DNA-Binding Proteins / metabolism
  • Doxorubicin / pharmacology*
  • Drug Interactions
  • Flow Cytometry
  • Histones / metabolism*
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Male
  • Phosphorylation / drug effects
  • Protein Kinase Inhibitors / pharmacology
  • Spermatozoa / cytology
  • Spermatozoa / drug effects*
  • Spermatozoa / metabolism*
  • Tumor Suppressor p53-Binding Protein 1
  • Wortmannin

Substances

  • Androstadienes
  • Antibiotics, Antineoplastic
  • DNA-Binding Proteins
  • H2AX protein, human
  • Histones
  • Intracellular Signaling Peptides and Proteins
  • Protein Kinase Inhibitors
  • TP53BP1 protein, human
  • Tumor Suppressor p53-Binding Protein 1
  • Doxorubicin
  • Acid Anhydride Hydrolases
  • RAD50 protein, human
  • DNA Repair Enzymes
  • Wortmannin