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Asian Journal of Andrology ; (6): 749-757, 2008.
Artigo em Inglês | WPRIM | ID: wpr-359914

RESUMO

<p><b>AIM</b>To investigate whether adriamycin induces DNA damage and the formation of gammaH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/repair proteins as somatic cells.</p>


Assuntos
Humanos , Masculino , Androstadienos , Farmacologia , Antibióticos Antineoplásicos , Farmacologia , Células Cultivadas , Ensaio Cometa , Quebras de DNA de Cadeia Dupla , Dano ao DNA , Enzimas Reparadoras do DNA , Metabolismo , Proteínas de Ligação a DNA , Metabolismo , Doxorrubicina , Farmacologia , Interações Medicamentosas , Citometria de Fluxo , Histonas , Metabolismo , Peptídeos e Proteínas de Sinalização Intracelular , Metabolismo , Fosforilação , Inibidores de Proteínas Quinases , Farmacologia , Espermatozoides , Biologia Celular , Metabolismo , Proteína 1 de Ligação à Proteína Supressora de Tumor p53
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