精子生成障碍患者Y染色体AZFc区部分缺失分析

目的:探讨人类Y染色体无精子症因子C区(AZFc)部分缺失对男性精子生成的影响。方法:选择Y染色体AZFc区9个序列标签位点(STS)sY1258、sY1291、sY254、sY255、sY1201、sY1206、sY1161、sY1197、sY1191,以ZFX/ZFY(X/Y连锁锌指蛋白基因)和SRY(sY14)基因为内对照。对160例Y染色体微缺失筛查均未发现缺失的无精子症及严重少精子症患者,76例正常生育男性DNA进行多重PCR扩增。疑有DAZ基因缺失的个体,采用基因单核苷酸变异分析(single nucleotide variants,SNV)技术,对DAZ基因4个拷贝中的单核苷酸多态位点进行检测,以确定DAZ基因的拷贝缺失类型。结果:160例无精子症及严重少精子症患者(病例组)gr/gr(sY1291)缺失10例,占6.3%;b2/b3(sY1191)缺失14例,占8.8%;新发现sY1291,sY1197缺失1例,占0.6%;b1/b2缺失1例,占0.6%;b1/b3缺失1例,占0.6%。76例正常生育男性(对照组)检出gr/gr缺失4例,占5.3%;b2/b3缺失4例,占5.3%。gr/gr缺失和b1/b3缺失(对照组和病例组)SNV分析均为DAZ1/DAZ2缺失;b2/b3缺失(对照组和病例组)SNV分析均为DAZ3/DAZ4缺失。1例sY1291,sY1197缺失的DAZ-SNVsY587位点缺失,1例b1/b2缺失者DAZ基因未缺失。结论:b2/b3(sY1191)缺失、gr/gr(sY1291)缺失在我国正常人群中多见,为基因组多态性;b1/b2缺失、b1/b3缺失和sY1291,sY1197缺失可能是导致精子生成障碍的高风险因子。

Partial AZFc region deletions of the Y chromosome in spermatogenic dysfunction patients

Objective:To investigate the influence of partial deletions in the AZFc region of the Y chromosome on spermatogenesis. Methods:We selected 9 sequence tagged sites (sY1258,sY1291,sY254,sY255,sY1201,sY1206,sY1161,sY1197 and sY1191) in the AZFc region of the Y chromosome,with ZFX/ZFY and SRY (sY14) as the interior control. We amplified by multiplex PCR the DNA of 160 patients with azoospermia or severe oligozoospermia that showed no microdeletion of the Y chromosome (the case group) and another 76 males with normal fertility (the control group). For the individuals suspected of DAZ gene deletion,we detected the single nucleotide polymorphism sites (SNPs) in the four copies of the DAZ gene by single nucleotide variation (SNV) analysis to determine the types of DAZ copy deletion. Results:In the case group,there were 10 cases of gr/gr (sY1291) deletion (6.3%),14 b2/b3 (sY1191) deletion (8.8%),1 sY1291,sY1197 deletion (0.6%),1 b1/b2 deletion (0.6%) and 1 b1/b3 deletion (0.6%),while in the control group,there were 4 cases of gr/gr deletion (5.3%) and 4 b2/b3 deletion (5.3%). SNV analysis showed DAZ1/DAZ2 deletion in all those with gr/gr and those with b1/b3 deletion,DAZ3/DAZ4 deletion in those with b2/b3 deletion,and DAZ-SNV sY587 deletion in 1 case of sY1291,sY1197 deletion,but no DAZ deletion was found in 1 case of b1/b2 deletion.Conclusion:b2/b3 (sY1191) and gr/gr (sY1291) deletions are genomic polymorphisms and quite common in the normal Chinese population; while b1/b2,b1/b3,and sY1291,sY1197 deletions may be high risk factors of dyszoospermia.