Department of Reproductive Biology and Stem Cells, Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.
Fertil Steril. 2013 Dec;100(6):1686-94.e1-7. doi: 10.1016/j.fertnstert.2013.07.1999. Epub 2013 Sep 4.
To identify potential biomarkers of azoospermia to determine a particular stage of spermatogenetic differentiation.
GeneChip Human Gene 1.0 ST microarray with validation at mRNA and protein levels.
Basic research laboratory.
PATIENT(S): Men with various types of nonobstructive azoospermia (n = 18) and with normal spermatogenesis (n = 4).
INTERVENTION(S): Obtaining 31 testicular biopsy samples.
MAIN OUTCOME MEASURE(S): Gene expression analysis using the Affymetrix Human Gene 1.0 ST microarrays on 14 selected genes according to the highest fold change, verified with quantitative polymerase chain reaction and on independent set of microarray samples. Western blot and immunohistochemistry were additionally performed.
RESULT(S): The comparative analysis of gene expression profiles in the infertile and control groups resulted in the selection of 4,946 differentially expressed genes. AKAP4, UBQLN3, CAPN11, GGN, SPACA4, SPATA3, and FAM71F1 were the most significantly down-regulated genes in infertile patients. Global analysis also led to identification of up-regulated genes-WBSCR28, ADCY10, TMEM225, SPATS1, FSCN3, GTSF1L, and GSG1-in men with late maturation arrest. Moreover, the results from quantitative polymerase chain reaction and Western blot largely confirmed the microarray data.
CONCLUSION(S): The set of selected genes can be used to create a molecular diagnostic tool to determine the degree of spermatogenic impairment for men with idiopathic nonobstructive azoospermia.
鉴定无精子症的潜在生物标志物，以确定生精分化的特定阶段。
在 mRNA 和蛋白质水平进行验证的 GeneChip Human Gene 1.0 ST 微阵列。
基础研究实验室。
各种类型的非阻塞性无精子症患者（n = 18）和正常生精患者（n = 4）。
获得 31 个睾丸活检样本。
根据最高倍数变化，使用 Affymetrix Human Gene 1.0 ST 微阵列对 14 个选定基因进行基因表达分析，并用定量聚合酶链反应和独立的微阵列样本进行验证。另外还进行了 Western blot 和免疫组织化学分析。
对不育组和对照组的基因表达谱进行比较分析，选择了 4946 个差异表达基因。AKAP4、UBQLN3、CAPN11、GGN、SPACA4、SPATA3 和 FAM71F1 是不育患者中表达下调最显著的基因。全局分析还导致鉴定出上调基因-WBSCR28、ADCY10、TMEM225、SPATS1、FSCN3、GTSF1L 和 GSG1-在晚期成熟阻滞的男性中。此外，定量聚合酶链反应和 Western blot 的结果在很大程度上证实了微阵列数据。
所选基因集可用于创建分子诊断工具，以确定特发性非阻塞性无精子症男性的生精损伤程度。
