The Expression Of VEGF-C,VEGFR-3 And NF-κ B P65 In Human Laryngeal Squamous Cell Carcinomas And Its Relationship With The Lympatic Nodes Metastesis

Objective: In order to promote early diagnosis and prevent lymphatic nodes metastasis of laryngeal carcinoma, to find a method to detect laryngeal carcinogenesis, we are study the relationship of the expression of vascular endothelial growth factor C(VEGF-C), vascular endothelial growth factor receptor 3 (VEGFR-3), neclear factor of kappa B(NF- κ B) and human laryngeal squamous cell carcinomas (LSCC), and compare with the expression of VEGF-C, VEGFR-3, NF- k B. To clarify the role of VEGF-C /VEGFR-3 in lymph nodes metastasis, and their relation with clinic pathlogic features, to promote diagnosis and therapy of LSCC. Methods: Protein levels of VEGF-C and NF- κ B expression were determined by immunohisto-chemistry in 60 cases of primary laryngeal carcinoma(PLC); The expression of VEGF-C and VEGFR-3 mRNA were examined in 30 cases of normal laryngeal mucosa tissue(NLM), PLC and cervical lymph node (CLN) by reverse transcription polymerase chain reaction (RT-PCR); And protein levelsof VEGF-C and VEGFR-3 expression were also determined by western blot in 20 cases of NLM, PLC, CLN, respectively. Results: Positive immunohistochemical staining for VEGF-C protein were observed in some laryngeal squamous carcinoma cells(45/60), the VEGF-C antigen was identified in the cytoplasm of cancer cells. Furthermore, the analysis show that the expression of VEGF-C mRNA and VEGFR-3 mRNA is different among NLM, PLC, CLN in the same patients. In PLC, expression of VEGF-C, VEGFR-3 was significantly higher in lymph node positive groups than in lymph node negative groups and associated with histological grade of differentiation (P<0.05); The expression of VEGF-C/VEGFR-3 were not associated with age, sex, site, T stage(P>0.05). There were overexpression of VEGF-C protein and VEGFR-3 protein in the CLN with western blot analysis, and associated with lymphatic nodes metastasis. Out of 60 cases of LSCC, 39 cases were found to have NF-κ B p65 protein positive stain, while none of the 10 cases of normal tissue was found to have NF- κ B p65 protein positive stain. The positive product was mainly localized in the cytoplasm of cancer cells. The expression rate of NF- κ B p65 protein in LSCC(65%) was significantly higher than that in normal tissue(P<0.01). The expression of NF-κ B p65 protein in LSCC were closely correlation with laryngeal canceration, but the expression of NF- κ B p65 protein was not related to the sex, age, location of carcinoma, lymph node metastasis and differentiation of LSCC and histological grade (P>0.05). Conclusion: These results suggest that there are a closely relationship in LSCC and VEGF-C /VEGFR-3, Namely, VEGF-C/VEGFR-3 could be an important factor regulating the mutual paracrine relationships between laryngeal cancer cells and endothelial cells inlymphatic vascular metastases, participated in regulating hyperplasia and permeability of lymphatic vessels and influencing metastatic fashion of cancer cells; But the expression of NF- k B p65 protein was not related with lymph nodes metastasis. So we are think that VEGF-C/VEGFR-3 probably have a prognostic significance in LSCC. Although NF- k B p65 help to diagnose early laryngocarcinoma, but it is not demonstrated that the expression of NF-k B p65 is a useful to judge prognosis of laryngocarcinoma.