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Asian Journal of Andrology ; (6): 749-757, 2008.
Artículo en Inglés | WPRIM | ID: wpr-359914

RESUMEN

<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>


Asunto(s)
Humanos , Masculino , Androstadienos , Farmacología , Antibióticos Antineoplásicos , Farmacología , Células Cultivadas , Ensayo Cometa , Roturas del ADN de Doble Cadena , Daño del ADN , Enzimas Reparadoras del ADN , Metabolismo , Proteínas de Unión al ADN , Metabolismo , Doxorrubicina , Farmacología , Interacciones Farmacológicas , Citometría de Flujo , Histonas , Metabolismo , Péptidos y Proteínas de Señalización Intracelular , Metabolismo , Fosforilación , Inhibidores de Proteínas Quinasas , Farmacología , Espermatozoides , Biología Celular , Metabolismo , Proteína 1 de Unión al Supresor Tumoral P53
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