| Object: To study the expression of vascular endothelial growth factor C(VEGF-C) and its receptor neuropilin-1(NRP-1) in laryngeal squamous cell carcinoma(LSCC) tissue and laryngocarcinoma Hep-2 cell lines, to discuss whether VEGF-C,NRP-1 have effect on proliferative metastasis facrors of laryngocarcinoma cells and if it can regulate the induction of LSCC cells on vascular formation in vitro.Methods: The expression of VEGF-C, NRP-1, mtP53, Bcl-2, Bax protein in 42 LSCC specimens and 20 polyp of cord specimens were detected by immunohistochemistry(IHC). Results of IHC staining were quantified by Image-Pro Plus analysis system. MTT method was applied to evaluate the effect of recombinate human VEGF-C and mouse NRP-1 antigen respectively on proliferation action of Hep-2 cells. IHC SP method, counting of migrated cells, adhesion, invasion and vascular formation in vitro experiment were applied to evaluate if there is any significant difference in the cell lines'migration, adhesion, invasion ability and cell lines'effect on vascular formation in vitro between the group treated with recombinate NRP-1 antigen and the control group.Result: All of the LSCC specimens were detected the expression of VEGF-C,NRP-1 protein, which was much higher than that in the polyp of cord tissue(P<0.05). The expression of VEGF-C,NRP-1 protein didn't significantly correlated with the clinical pathological factors such as gender, age(P>0.05) but with tumor T stage and lymph node metastasis(P<0.05). The expression of NRP-1 correlated with grade of tumor cell differentiation(P<0.05), but VEGF-C didn't significantly correlated with grade of tumor cell differentiation(P>0.05). There was no significant correlation between the expression of VEGF-C and mtP53 protein, Bcl-2 protein, Bax protein, Bcl-2/Bax rate(r=0.189 P>0.05, r=0.264 P>0.05,r=0.092 P>0.05, r=0.130 P>0.05). The expression of NRP-1 protein didn't correlate with Bax protein(r=0.129 P>0.05) but significantly with mtP53 protein, Bcl-2 protein, Bcl-2/Bax rate(r=0.606 P<0.01, r=0.842 P<0.01, r=0.605 P<0.01). The Hep-2 cell lines expressed both VEGF-C and NRP-1,VEGF-C had no significant positive influence on proliferantion of Hep-2(P>0.05). The proliferation inhibition rate of Hep-2 cells rose up as the recombination NRP-1 antigen's concentration increaced(r=0.624 P<0.01). In the group of NRP-1 antigen, the expression of mtP53, Bcl-2 protein was much less than that in the control group evaluated by cell IHC(P<0.01). The number of cells migration in the group of NRP-1 antigen was fewer than that in the control group(P<0.01). After the Hep-2 cells treated with NRP-1 antigen for 48 hours, compared with the control group, the adhesion, invasion ability was significantly weakened(P<0.01) and induction of vascular lumina on matrigel in vitro was much less (P<0.01).Conclution:1. VEGF-C and NRP-1 proteins over-express in LSCC and they correlate with T stage and regional lymph node metastasis,suggest that VEGF-C and NRP-1 play an important role in development and metastasis of LSCC.2. NRP-1 antigen has significant effect on inhibition of Hep-2 cell lines, It may inhibit cell apoptosis through down-regulating the mtP53,Bcl-2 expression; NRP-1 antigen has significant inhibitive effect on the migration, adhesion and invasion ability of Hep-2 cell lines, and could weaken the vascular formation induction of Hep-2 cells in vitro.3. VEGF-C has no significant effect on proliferation of Hep-2 cell lines.4. In laryngocarcinoma Hep-2 cell lines,NRP-1 is not a independent receptor to VEGF-C.
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