Open AccessAdriamycin induces H2AX phosphorylation in human spermatozoa
Author(s) -
Li ZhongXiang,
Wang TingTing,
Wu YanTing,
Xu ChenMing,
Dong MinYue,
Sheng JianZhong,
Huang HeFeng
Publication year - 2008
Publication title -
asian journal of andrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.701
H-Index - 74
eISSN - 1745-7262
pISSN - 1008-682X
DOI - 10.1111/j.1745-7262.2008.00400.x
Subject(s) - comet assay , wortmannin , flow cytometry , dna damage , histone , microbiology and biotechnology , biology , phosphorylation , staining , dna repair , chemistry , dna , phosphatidylinositol , biochemistry , genetics
Aim: To investigate whether adriamycin induces DNA damage and the formation of γH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa. Methods: Human spermatozoa were treated with adriamycin at different concentrations. γH2AX 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 μg/mL for different times (0.5, 2, 8 and 24 h), or for 8 h at different concentrations (0.4, 2 and 10 μg/mL), induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of γH2AX 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 γH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3‐kinase (PI3K) family, abolished the co‐appearance of these two proteins with γH2AX. Conclusion: Human mature spermatozoa have the same response to DSB‐induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/ repair proteins as somatic cells.