Adriamycin induces H2AX phosphorylation in human spermatozoa.
- Author:
Zhong-Xiang LI
1
;
Ting-Ting WANG
;
Yan-Ting WU
;
Chen-Ming XU
;
Min-Yue DONG
;
Jian-Zhong SHENG
;
He-Feng HUANG
Author Information
- Publication Type:Journal Article
- MeSH: 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; drug effects; metabolism; Tumor Suppressor p53-Binding Protein 1
- From: Asian Journal of Andrology 2008;10(5):749-757
- CountryChina
- Language:English
-
Abstract:
AIMTo investigate whether adriamycin induces DNA damage and the formation of gammaH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa.
METHODSHuman 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.
RESULTSThe 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.
CONCLUSIONHuman mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/repair proteins as somatic cells.
