Neuropilin-1 promotes HepG2 proliferation, invasion and migration through potentiating the activity of hepatic stellate cells

Abstract

Objective To explore the effect of neuropilin-1 (NRP-1) on tumor cells proliferation,invasion and migration through activation of hepatic stellate cells in primary liver cancer.Methods An adenovirus vector containing human NRP-1 gene (pDC315-NRP-1-3Flag) was constructed at first,LX2 cells was infected with the adenovirus vector,the expression of NRP-1 in the LX2 was observed by Western blotting,the expression of α-smooth muscle actin (α-SMA) and NRP-1 in LX2 was observed by immunohistochemical staining.LX2 and HepG2 were cocultured,the proliferatio of LX2 and HepG2 were observed by methyl thiazol tetrazolium (MTT),the invasion and migration of HepG2 was observed by Transwell chamber.Results The adenovirus vector containing human NRP-1 gene (pDC315-NRP-1-3Flag) was constructed successfully,the best virus titer for cell infection was 2.0 × 106 PFU/ml and the infection efficiency was above 90％ after 48 h.Western blotting showed the expression of NRP-1 was higher in the transfection group than in the mock-vehicle and non-transfection groups,and the expression of NRP-1 was also higher after infected for 48 h than for 24 h.Immunohistochemical results showed that the expression of α-SMA and NRP-1 were higher in the experimental group than in the mock-vehicle and non-transfection groups.MTT results showed that the proliferation of LX2 was higher in the experimental group than in the two latter groups (P ＜ 0.05),the difference was significant.Coculture experiments showed that LX2 could promote proliferation,invasion and migration of HepG2,the promoting effect in the experimental group was significant higher than in the control groups (P ＜ 0.05).Conclusion LX2 cells could be infected with constructed adenovirus vector containing human NRP-1 gene stably.NRP-1 could promote activion and proliferation of LX2,which in turn promotes proliferation,migration and invasion of HepG2 cells. Key words: Neuropilin-1 ; Primary liver cancer; Hepatic stellate cell; Tumor microenvironment