Hypospadias is a common congenital abnormality of the penile. Abnormal regulation of critical genes involved in urethral development leads to hypospadias. We used the Rab25 mice and foreskin fibroblasts transfected with lentivirus in vitro and in vivo to investigate the role of Rab25 in hypospadias. The expression levels of various molecules in tissue samples and foreskin fibroblasts were confirmed using molecular biology methods (western blotting, PCR, immunohistochemistry, etc.). A scanning electron microscope (SEM) was used to visualize the external morphology of genital tubercles (GTs) of gestation day (GD) 18.5 male wild-type (WT) and Rab25 mice. An expanded distal cleft and V-shaped urethral opening were observed in GD 18.5 Rab25 mice. We demonstrated that Rab25 mediated hypospadias through the β1 integrin/EGFR pathway. In addition, silencing Rab25 inhibited cell proliferation and migration and promoted apoptosis in the foreskin fibroblasts; Ki-67- and TUNEL-positive cells were mainly concentrated near the urethral seam. These findings suggest that Rab25 plays an essential role in hypospadias by activation of β1 integrin/EGFR pathway, and Rab25 is a critical mediator of urethral seam formation in GD18.5 male fetal mice.

中文翻译：

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Rab25 通过 β1 整合素/EGFR 途径参与尿道下裂

尿道下裂是一种常见的阴茎先天性异常。参与尿道发育的关键基因的异常调节会导致尿道下裂。我们利用慢病毒转染的Rab25小鼠和包皮成纤维细胞在体外和体内研究Rab25在尿道下裂中的作用。采用分子生物学方法（蛋白质印迹、PCR、免疫组化等）证实组织样本和包皮成纤维细胞中各种分子的表达水平。使用扫描电子显微镜 (SEM) 观察妊娠日 (GD) 18.5 雄性野生型 (WT) 和 Rab25 小鼠生殖器结节 (GT) 的外部形态。在 GD 18.5 Rab25 小鼠中观察到扩大的远端裂口和 V 形尿道开口。我们证明 Rab25 通过 β1 整合素/EGFR 途径介导尿道下裂。此外，沉默Rab25可抑制细胞增殖和迁移，并促进包皮成纤维细胞凋亡。 Ki-67和TUNEL阳性细胞主要集中在尿道缝附近。这些发现表明，Rab25 通过激活 β1 整合素/EGFR 通路在尿道下裂中发挥重要作用，并且 Rab25 是 GD18.5 雄性胎鼠尿道缝形成的关键介质。