精子发生或成熟异常是男性不育的最主要原因。研究表明，非编码 RNA如miRNA、长非编码 RNA（lncRNA）在精子发生过程中起着重要作用。申请人前期发现，lncRNA NLC1-C在生精细胞成熟停滞患者（MA）睾丸组织中表达下调，且聚集在细胞核中；NLC1-C结合蛋白Nucleolin在MA中表达亦下降，其表达定位在正常睾丸组织的初级精母细胞核和MA的精原细胞核中。这些结果提示NLC1-C在男性不育中发挥重要作用。但NLC1-C作用的确切分子机制有待进一步探讨，因此，本项目拟开展：1）检测NLC1-C及Nucleolin蛋白在不同类型男性不育睾丸组织中表达情况；2）分别对NLC1-C/Nucleolin表达紊乱的病理组织和体外模拟的睾丸癌细胞进行高通量基因测序，探究因NLC1-C/Nucleolin蛋白表达异常导致MA的相关基因，并探讨其分子机制,为男性不育的诊治提供新的线索。

Spermatogenic failure is one of the major reasons of male infertility. Non-coding RNAs, eg. microRNAs and long non-coding RNAs, have been recently emerged as key regulators of eukaryotic gene expression in the spermatogenesis. Our previous results indicated that long non-coding RNA NLC1-C was predominately expressed in both cytoplasm and nuclei of spermatogonia and primary spermatocytes in spermatogenesis, but cytoplasmic NLC1-C was significantly down-regulated and nuclear NLC1-C was increased in male infertility patients with mixed pattern of maturation arrest (MA). NLC1-C associated protein Nucleolin was mainly localized in spermatocytes, but in testicular samples of MA, the spermatocytes signal was excluded and a dislocated expression of Nucleolin was observed in spermatogonia. Our results demonstrated that NLC1-C was a key regulator in spermatogenesis failure. However, the role and the molecular mechanism of NLC1-C in MA need to be further investigated. Thus, our project is aimed to: 1) detect the expression of NLC1-C and Nucleolin in different types of non-obstructive azoospermia (NOA), including MA and hypospermatogenesis patients’ testicular biopsies; 2) investigate the molecular mechanism of NLC1-C and its associated protein Nucleolin in MA. To achieve this goal, I will use high throughput sequencing to identify and characterize genes involved in NLC1-C/Nucleolin pathway in spermatogenesis. Our study will provide new clues to personalized diagnosis and treatment for male infertility.