MiR-378a-5p inhibits angiogenesis of oral squamous cell carcinoma by targeting KLK4

 Zhi Cui, Qi-Lin Liu, Shi-Qun Sun, Kun Jiao, Dong-Rui Liu, Xue-Chun Zhou, Lei Huang

Abstract:

Oral squamous cell carcinoma (OSCC) is still a leading cause of cancer death owing to distant metastasis, which is largely facilitated by tumor angiogenesis. MicroRNA (miR)-378a-5p and Kallikrein-related peptidase 4 (KLK4) participate in tumorigenesis and tumor metastasis according to previous studies, yet the exact role they play in tumor angiogenesis remains poorly understood. The aim of the present study was to investigate the effect of miR-378a-5p and KLK4 on angiogenesis of OSCC. MTT assay showed that the expression level of miR-378a-5p was negatively correlated with the proliferation of OSCC cells. ELISA and western blot assay showed that downregulation of miR-378a-5p promoted VEGF expression. Tube formation and in vivo chicken chorioallantoic membrane (CAM) assay showed that inhibition of miR-378a-5p reduced tube formation of human umbilical vein endothelial cells (HUVECs) and newly formed microvessel. On the contrary, overexpression of KLK4 enhanced angiogenesis of OSCC cells with increased VEGF expression, tube formation activity of HUVECs and newly formed microvessel. Moreover, the dual-luciferase assay validated that KLK4 was a target gene of miR-378a-5p. MiR-378a-5p silencing induced tube formation was suppressed by the downregulation of KLK4. Besides, the activation of Wnt/β-catenin signaling pathway in miR-378a-5p antagomir transfected cells was also blocked by the KLK4 shRNA. To sum up, our study suggests that miR-378a-5p suppressed angiogenesis of OSCC at least partly by the regulation of KLK4.