Hypospadias is an abnormal ventral development of the penis caused by incomplete virilization of the male genital tubercle. This study investigated the phenotypic modulation of vascular smooth muscle cells (VSMCs) in the corpus spongiosum surrounding the urethral plate in hypospadias. The urethral corpus spongiosum tissue was collected for HE, Masson and α‐SMA immunohistochemical staining. Spongiosum VSMCs were cultured and identified by α‐SMA fluorescence. qRT‐PCR and Western blotting and fluorescence were performed. The results showed that the vascular lumen of the corpus spongiosum around the urethral plate was larger and that the vascular smooth muscle layer was thicker in hypospadias. The expression of the contractile markers α‐SMA and Calponin 1 in VSMCs was decreased, the expression of the synthetic marker OPN was increased, and the transcription of the phenotypic switching factors SRF and MYOCD was decreased. The expression of Ki67, PCNA and BAX was increased, and the expression of Bcl‐2 was decreased. The phenotype of corpus spongiosum VSMCs in hypospadias changed from the contractional type to the synthetic type. This phenotypic modulation was associated with increased proliferation and apoptosis rates. SRF and MYOCD may be the main factors mediating the phenotypic modulation of urethral corpus spongiosum VSMCs.
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