Animal model studies have shown, MSY2 gene has a potential role in spermatogenesis. Some mutations on this gene have been proposed to be associated with human male infertility. In this study, polymorphisms of exon1 of YBX2 gene have been investigated. A total of 156 men were evaluated. They included 96 men with normal spermatogenesis, 60 men with non-obstructive azoospermia. We extracted DNA from blood and testis tissues of samples, and analyzed polymorphisms of exon1 by sequencing method. Sequencing results showed that among the studied polymorphisms, frequency of TT genotype in rs222859 polymorphism was significantly higher in azoospermic patients compared to control group (P<0.001). According to our results, YBX2 gene may play a role in male reproduction. The alterations of this gene might be involved in azoospermia among the Iranian population.