NCK1 promotes the angiogenesis of cervical squamous carcinoma via Rac1/PAK1/MMP2 signal pathway

• Published in: Gynecologic oncology 2019-02, Vol.152 (2), p.387-395
• Main Authors: Xia, Pei, Huang, Mingchuan, Zhang, Yuting, Xiong, Xiujuan, Yan, Min, Xiong, Xiaoliang, Yu, Weiwei, Song, Enlin
• Format: Article
• Language: English
• Subjects: Adaptor Proteins, Signal Transducing - biosynthesis; Adaptor Proteins, Signal Transducing - genetics; Adaptor Proteins, Signal Transducing - metabolism; Angiogenesis; Carcinoma, Squamous Cell - blood supply; Carcinoma, Squamous Cell - genetics; Carcinoma, Squamous Cell - metabolism; Cell Line, Tumor; CSCC; Female; Hematology, Oncology, and Palliative Medicine; Human Umbilical Vein Endothelial Cells; Humans; Matrix Metalloproteinase 2 - metabolism; Middle Aged; MMP2; Nck1; Neovascularization, Pathologic - genetics; Neovascularization, Pathologic - metabolism; Neovascularization, Pathologic - pathology; Obstetrics and Gynecology; Oncogene Proteins - biosynthesis; Oncogene Proteins - genetics; Oncogene Proteins - metabolism; p21-Activated Kinases - metabolism; PAK1; Rac1; rac1 GTP-Binding Protein - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Signal Transduction; Up-Regulation; Uterine Cervical Neoplasms - blood supply; Uterine Cervical Neoplasms - genetics; Uterine Cervical Neoplasms - metabolism
• ISSN: 0090-8258; 1095-6859
• DOI: 10.1016/j.ygyno.2018.11.013

AbstractObjectiveThe study was to explore the roles of Nck1 in the angiogenesis of cervical squamous cell carcinoma (CSCC). MethodsmRNA and protein levels were evaluated with real-time quantitative PCR and immunohistochemisty/western blotting respectively. The cancer microvessel density (MVD) was assayed with CD34 endothelial labeling. Nck1 gene knock-in (SiHa-Nck1+) and knock-down (SiHa-Nck1-) were achieved by gene transfection and siRNA respectively. Protein level from cellular supernatant was measured with ELISA. Proliferation, migration and tube formation of the Human Umbilical Vein Endothelial cells (HUVECs) were evaluated by CCK-8 cell viability assay, transwell chamber assay and in vitro Matrigel tubulation assay respectively. ResultsNck1 level gradually increased from normal cervical epithelia to high-grade CIN, overexpressed in CSCC and was associated with cancer MVD. The ability of proliferation, migration and tube formation of HUVECs was enhanced in SiHa-Nck1+-treated while decreased in SiHa-NcK1−-treated cells compared to SiHa-control-treated cells. Mechanistically, RAC1-GTP, p-PAK1 and MMP2 were increased in SiHa-NCK1+ cells and pretreatment with the Rac1 inhibitor (NSC23766) significantly decreased their levels. Furthermore, inhibition of PAK1 reduced MMP2 level in SiHa-Nck1+ cells whereas the level of Rac1-GTP was unaltered. Also, inhibition of Rac1 or PAK1 impaired angiogenesis-inducing capacity of cancer cells. ConclusionsNck1 promotes the angiogenesis-inducing capacity of CSCC via the Rac1/PAK1/MMP2 signal pathway.