The influence of NRP-1 gene on the migration, invasion and angiogenesis of human bladder urothel carcinoma cells in vitro

Abstract

Objective To determine the best targets that can effectively interfere NRP-1 for the research of NRP-1 functional gene and explore the invasion, transfer and ability to promote nearby angiogenesis of NRP-1 by employing lentiviral to interfere with NRP-1 gene expression in two bladder urothelial cell carcinoma cell lines, so as to provide a theoretical basis for the pathogenesis and transfer mechanism of bladder urothelial cell carcinoma. Methods The Western Blot was adopted to detect the NRP-1 protein expression levels to verify two bladder urothelial cell carcinoma cell lines as the follow-up test models. RNA interference targeting NRP-1 were designed. Interference vectors constructed and packaged in lentiviral vectors were infected with above two cells, and the best target to effectively interfere NRP-1 was identified by the qPCR and Western Blot. The two bladder cancer cell lines were respectively infected with controlled lentivirus and interfering NRP-1 gene lentiviral vector, to verify the invasion, transfer and ability to promote nearby angiogenesis of NRP-1 in bladder urothelial cell carcinoma cells by Transwell method and angiogenesis method. Results By using the Transwell smallchamber, it was found that in-vitro invasion and migration capabilities of two bladder urothelial cell carcinoma cell lines infected with control lentiviral virus were significantly higher than the cells infected with NRP-1 interference lentiviral virus. The in-vitro invasion and migration inhibition rates of NRP-1 interference on T24 was 51% and 72%, respectively (P<0.05), while those on 5637 was 61% and 65%, respectively (P<0.05). It was found through angiogenesis experiments that compared to the control group(virus-infected), the two bladder urothelial cell carcinoma cell lines in the NRP-1 interference group had declined influences on the angiogenesis ability of vascular endothelial cells. In the interference group, the ability to control angiogenesis decreased by 21% and 28% in T24 and 5637, respectively (P<0.05). Conclusions NRP-1 lentiviral RNA interference inhibits the migration and invasion of bladder urothelial cell carcinoma cells.NRP-1 interference has inhibitory effect on angiogenesis around bladder urothelial cell carcinoma cells. Key words: Urinary Bladder Neoplasms; Neuropilin-1; Neoplasm Invasiveness