Department of Surgery, The Affiliated Huaian No. 1 People's Hospital of Nanjing Medical University, Huaian, Jiangsu, China.
State Key Laboratory of Reproductive Medicine, Center for Global Health, Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing, Jiangsu, China.
Mol Genet Genomic Med. 2019 Sep;7(9):e870. doi: 10.1002/mgg3.870. Epub 2019 Jul 21.
It has been proposed that lncRNAs, widely transcribed from genomes, play pivotal regulatory roles in a variety of biological processes, but their function in regulating spermatogenesis in human males is rarely reported.
QRT-PCR was adopted to detect HOTTIP expression level in testicular tissues from hypospermatogenesis (Hypo) patients or controls. The proliferation levels of NT2 and 293T were measured via CCK-8 and EdU detection. Meanwhile, luciferase reporter gene assay and bioinformatics analysis were carried out to identify a target of HOTTIP. Additionally, the underlying mechanism of HOTTIP's function was investigated using western blotting and RIP analysis.
The research results manifested that the expression of HOTTIP in testicular tissues from Hypo patients was prominently reduced in comparison with that in control testicular tissues. Interestingly, it was noted that HOTTIP exhibited a high expression in testicular embryonal carcinoma cell line NT2 compared with that in normal control cell line 293T. It was denoted in cell function evaluation that cell proliferation was impeded by downregulated HOTTIP but evidently stimulated by overexpressed HOTTIP. Moreover, HOTTIP was capable of positively modulating HOXA13 expression via the competitive binding to miR-128-3p.
Therefore, HOTTIP acting as ceRNAs to promote testicular embryonal carcinoma cell proliferation.
长链非编码 RNA（lncRNAs）广泛转录于基因组中，被认为在多种生物学过程中发挥关键的调控作用，但它们在调控人类男性精子发生中的功能却鲜有报道。
采用 QRT-PCR 检测少精子症（Hypo）患者或对照睾丸组织中 HOTTIP 的表达水平。通过 CCK-8 和 EdU 检测来测量 NT2 和 293T 的增殖水平。同时，进行荧光素酶报告基因检测和生物信息学分析，以鉴定 HOTTIP 的靶标。此外，通过 Western blot 和 RIP 分析研究 HOTTIP 功能的潜在机制。
研究结果表明，Hypo 患者睾丸组织中的 HOTTIP 表达明显低于对照组睾丸组织。有趣的是，研究发现 HOTTIP 在睾丸胚胎癌细胞系 NT2 中的表达明显高于正常对照细胞系 293T。细胞功能评估表明，下调 HOTTIP 会抑制细胞增殖，但过表达 HOTTIP 则会明显刺激细胞增殖。此外，HOTTIP 通过与 miR-128-3p 的竞争性结合正向调控 HOXA13 的表达。
因此，HOTTIP 作为 ceRNA 促进睾丸胚胎癌细胞增殖。
