ABSTRACT
Objective To investigate whether γH2AX could be a useful biomarker for evaluating the DNA double‐stranded . Methods Semem samples in case group were from 27 infertile males who were diagnosed in Andriatrics department or reproductive centre in the First Affiliated Hospital of Guangxi Medical University .The other semen samples were from 23 healthy donors with fertility as comparison .The levels of γH2AX were detected by flow cytometry .Single cell gel electropherosis(SCGE)was applied to assess the level of DSBs of sperm .Density gradient centrifugation(DGC) was applied to optimized spermatozoa .Results TheγH2AX levels and the DSBs of the sperm of the infertile subjects were significantly higher than those of healthy males(P<0 .01) , and the levels of γH2AX and the DSBs of sperm significantly decreased in two groups by DGC(P<0 .01) .Conclusion The level of spermatozoaγH2AX is higher in male infertility patients than in healthy donors with fertility ,which might be a useful biomarker for evaluating DSBs of sperm .
ABSTRACT
Objective To discover the role of MCPH1 in DNA double-strand damage induced by ionizing radiation and its relationship with H2AX in esophageal cancer cell ECA109. Methods ECA109 cancer cells accepted 8 Gy 1 h after irradiation were collected for protein extraction and immunofluorescence then MCPH1 and H2AX protein expression and nuclear foci changes were observed. A stable low expression of H2AX cell lines was established and MCPH1 and H2AX protein expression and nuclear foci changes induced by ionizing radiation after silence H2AX were detected. Results (1)A stable low expression of H2AX cell lines in ECA109 cells was successfully constructed. (2)Ionizing radiation could cause the increase of r-H2AX and MCPH1 protein expression, as the same as nuclear focus increase of r-H2AX and MCPH1. (3)The protein level and nucleus focus of r-H2AX and MCPH1 were significantly reduced in ECA109 after silence H2AX. Conclusion MCPH1 is the part of DNA damage response triggered by ionizing radiation and is located in damage response downstream and can be regulated by H2AX.