The Roles of Differentially Expressed Genes in Human Cryptorchid Testes on testicular differentiation and development

目的：为了恢复生育能力，目前的共识建议早期睾丸固定术治疗隐睾症。尽管早期睾丸固定术，然而，据报道，转化成成年深色精原细胞的生殖细胞已经受损，在操作时，因此影响未来的生育能力。为了阐明隐睾生殖细胞成熟过程中发生的生物学过程，我们确定了受睾丸发育不良影响的基因，采用基于PCR的抑制性消减杂交，并调查差异表达的基因，以确定它们与细胞分化和精子发生。材料与方法：24例12-59个月的男性睾丸标本，均行睾丸固定术或鞘膜积液切除术。取同侧隐睾和睾丸下降的年龄匹配的男性睾丸组织，进行双向减影。通过实时荧光定量RT-PCR验证差异表达，并通过免疫组化和western blot分析候选基因的分布。结果：我们获得了84个克隆，对应于代表差异表达的转录本。BLAST检索发现32个不同的已知基因，相似性为98-100%。其中，我们进一步研究了三个基因，TPT 1，EEF 1A 1和NuMA 1，这是显着更高的表达在隐睾比在下降的睾丸，并检测到精原细胞从未成熟到成年睾丸。结论：TPT 1、EEF 1A 1和NuMA 1具有与细胞生长相关的功能，提示它们在生殖细胞分化和维持干细胞潜能中发挥一定作用。这些基因在睾丸中表达水平的变化可能使隐睾引起的生精障碍的新的评估。

Purpose : In order to restore fertility, current consensus recommends early orchiopexy for cryptorchidism. Despite early rchiopexy, however, it is reported that transformation of gonocytes into adult dark spermatogonia is already impaired at the time of operation and consequently affects future fertility. To elucidate the biological processes occurring during germ cell maturation in the cryptorchid testes, we identified the genes affected by testicular maldescent using PCR-based suppression subtractive hybridization, and investigated differentially expressed genes in order to determine they are related to cell differentiation and spermatogenesis. Materials and Methods : Testicular tissues were excised from 24 boys who underwent orchiopexy or hydrocelectomy in our hospital (12-59 months old). Between their tissues from age-matched boys with ipsilateral cryptorchidism and descended testis, two-way subtraction was performed. Differential expression was validated by real-time RT-PCR.Furthermore, to clarify the distribution of candidate genes, immunohistochemistry and western blot analysis were performed. Results: We obtained 84 clones corresponding to transcripts representing differential expression. BLAST searches revealed 32 different known genes with 98-100% similarity. Among these, we further investigate three genes, TPT1, EEF1A1 and NuMA1, that were significantly more highly expressed in cryptorchidism than in descended testes, and were detected in the spermatogonia from immature to adult testes. Conclusions : TPT1, EEF1A1 and NuMA1 have cell growth-related functions, suggesting that they play certain roles in germ cell differentiation and maintenance of stem cell potential. Changes in the expression levels of these genes expressions in the testes might enable novel evaluation of spermatogenic failure caused by cryptorchidism.