Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation
Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation
复制标题
成纤维细胞生长因子 5 通过 ERK 和 AKT 激活促进精原干细胞增殖
DOI：
10.1186/s13287-019-1139-7
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发表时间：
2019-01-22
影响因子：
7.5
通讯作者：
Li, Zheng
中科院分区：
文献类型：
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作者：
Tian, Ruhui;Yao, Chencheng;Li, Zheng
支持细胞是构成精原干细胞微环境（称为生态位）最重要的体细胞。它们产生大量的关键生长因子和结构蛋白，在雄性ssc的存活、增殖和分化的复杂过程中发挥重要作用。有人认为，支持细胞异常可能导致精子发生失败，最终导致人类无精子症。然而，到目前为止，人类支持细胞的基因表达特征和蛋白质功能仍不清楚。在这项研究中，我们旨在评估成纤维细胞生长因子-5 （FGF5）对SSCs的影响。FGF5是一种新型生长因子，在支持细胞综合征（SCOS）患者的支持细胞中下调，而在阻塞性无精子症（OA）患者的支持细胞中下调。方法比较SCOS和OA患者支持细胞的转录组。然后，我们评估了FGF5（一种在SCOS支持细胞中下调的生长因子）在人原代培养的支持细胞和睾丸组织中的表达。采用EDU法和CCK-8法检测FGF5在小鼠ssc中的增殖作用。为了研究FGF5的作用机制，我们使用了Phospho Explorer Array。Western blot法对结果进行验证。结果通过RNA-Seq分析，我们发现SCOS和OA患者的支持细胞存在308个差异表达基因（DEGs）。我们注意到并证实，在转录和翻译水平上，OA患者的支持细胞中成纤维细胞生长因子-5 （FGF5）的表达高于SCOS患者。增殖实验显示，rFGF5对小鼠ssc细胞系C18-4的增殖作用呈时间和剂量依赖性。此外，我们发现rFGF5激活了ERK和AKT，并增强了Cyclin A2和Cyclin E1的表达。结论采用RNA- seq技术鉴定出SCOS和OA患者的支持细胞具有明显的RNA谱。FGF5是一种在SCOS支持细胞中下调的生长因子，可通过激活ERK和AKT促进SSCs增殖。
BackgroundSertoli cells are the most important somatic cells contributing to the microenvironment (named niche) for spermatogonial stem cells (SSCs). They produce amounts of crucial growth factors and structure proteins that play essential roles in the complex processes of male SSCs survival, proliferation, and differentiation. It has been suggested that Sertoli cell abnormalities could result in spermatogenesis failure, eventually causing azoospermia in humans. However, to the end, the gene expression characteristics and protein functions of human Sertoli cells remained unknown. In this study, we aimed to evaluate the effect of fibroblast growth factor-5 (FGF5), a novel growth factor downregulated in Sertoli cells from Sertoli cell-only syndrome (SCOS) patients compared to Sertoli cells from obstructive azoospermia (OA) patients, on SSCs.MethodsWe compared the transcriptome between Sertoli cell from SCOS and OA patients. Then, we evaluated the expression of FGF5, a growth factor which is downregulated in SCOS Sertoli cells, in human primary cultured Sertoli cells and testicular tissue. Also, the proliferation effect of FGF5 in mice SSCs was detected using EDU assay and CCK-8 assay. To investigate the mechanism of FGF5, Phospho Explorer Array was performed. And the results were verified using Western blot assay.ResultsUsing RNA-Seq, we found 308 differentially expressed genes (DEGs) between Sertoli cells from SCOS and OA patients. We noted and verified that the expression of fibroblast growth factor-5 (FGF5) was higher in Sertoli cells of OA patients than that of SCOS patients at both transcriptional and translational levels. Proliferation assays showed that rFGF5 enhanced the proliferation of mouse SSCs line C18-4 in a time- and dose-dependent manner. Moreover, we demonstrated that ERK and AKT were activated and the expression of Cyclin A2 and Cyclin E1 was enhanced by rFGF5.ConclusionThe distinct RNA profiles between Sertoli cells from SCOS and OA patients were identified using RNA-Seq. Also, FGF5, a growth factor that downregulated in SCOS Sertoli cells, could promote SSCs proliferation via ERK and AKT activation.