Research On The Role Of Vegf-d And Vegfr-3 In Lymph Metastasis In The Squamous Cell Carcinoma Of Tongue

BackgroundThe squamous cell carcinoma of tongue was one common malignant tumor of oral maxillo-facial region.It had such characteristics of high degree malignancy,rapid growth,invasion and high rate of lymph metastasis et al.The ratio of lymph metastasis was about 32.3%in our nation.It was reported about between 21%-52%in Europe and America.Studies reported that when the size of the squamous cell carcinoma of tongue was in situ,new vessels would be generated to provide essential oxygen and nutrition under the control effect of many positive or negative factors.It was deficient of study of the mechanism of dilatation of lymphatic vessel and relationship of the lymphatic vessel and lymphatic metastasis.Vascular endothelial growth factor D(VEGF-D) was a member of the VEGF family,and it was a lymph angiogenesis promoting factor.It was different in the study results of the expression of VEGF-D.VEGF-D could bind with its receptor VEGFR-3, induce lymph metastasis and dilation of lymphatic vessels in tumor tissue.It was to be confirmed of the relationship with VEGF-D\VEGFR-3 and lymph metastasis.In the Past,lymph angiogenesis study had been limited because that the lacking of specific lymphatic endothelial markers maked it difficult to distinguish the blood vessel and the lymphatic vessel on morphology only by HE staining technique.It was reported by Marks in 1999 that D2-40 was a monoclonal antibody that reacts with a 40-kd onco fetal antigen,better known as M2A antigen,that was expressed in fetal testis and germ cell.The new studies reported that D2-40 was usually reported to detect lymphatic vessels selectively in the tissues of gastric cancer,prostate cancer and malignant melanoma by reacting selectively with the fixed antigen on lymphatic endothelium without reaction with mature lymphatic vessel and capillary vessel endothelium.D2-40 had been a specific marker of lymphatic vessel.Objective:To detect the expressive levels of VEGF-D(vascular endotbelial growth factor-D) and VEGFR-3 in squamous cell carcinoma of tongue.To determine of the density of lymphatic vessels marked by D2-40.To analyse the correlation of the density of lymphatic vessel and the family of VEGF with lymph metastasis in the squamous cell carcinoma of tongue,and the correlation with its clinicoPathological factors in squamous cell carcinoma of tongue.To conclude the possible role of VEGF-D\VEGFR-3\LVD of lymph metastasis in squamous cell carcinoma of tongue.Materials and methods69 specimens of squamous cell carcinoma of tongue were gathered from the Department of Pathology,the Fist Affiliated Hospital of Zhengzhou University from July 2005 to 2006(All the cases were without Preoperative chemotheraphy or radiotherapy).20 specimens of normal tongue mucosa tissues were served as control group.The specimens were all fixed with 10%formalin and embedded with paraffin routinely.And each tissue block was stained with hematoxylin and eosin for histopathological analysis and the others were used for immunostaining. Immunohistochemistry method was applied to determine the expression of VEGF-D,VEGFR-3 and D2-40 in each sample.The data were analyzed by statistics software SPSS13.0.Chi-squared test orrelation were performed to compare different between groups and correlated analysis respectively.The significant difference was considered when the P value was less than 0.05.The student t-test and F-test were used to analyze data.Results:1,Expression of VEGF-D in the normal tongue mucosa tissues was less and observed in the cytoplasm.The positive rates was 15.00%.Expression of VEGF-D in the squamous cell carcinoma of tongue was observed in the cytoplasm of tumor.The positive rates was 43.48%.There were significant differences in both of them and normal tissues(p＜0.05).The positive rates was 59.46%in the group of lymph node metastasis.The positive rates was 25.00%in the group of non lymph node metastasis. There were significant differences in both of them(p＜0.05).No correlation was found with age,tumor diameter,histologic types(p＜0.05).2,Expression of VEGFR-3 in the squamous cell carcinoma of tongue was mainly observed in the endothelial cell of lymphatic vessel of tumor and also observed in the cytoplasm of tumor.The positive rates was 63.77%.There were significant differences in both of them and normal tissues(p＜0.05).The positive expression of VEGF-D in the squamous cell carcinoma of tongue showed obvious correlation with the positive expression of VEGFR-3(p＜0.05) and obvious correlation with lymph metastasis(p＜0.05).3,The D2-40 marked positive lymphatic vessels which had thin wall,irregular lumens and shapes were in interstitial tissue.There was less in the normal tissues. LVD value(33.24±0.09) in the squamous cell carcinoma of tongue was higher than in the normal tongue mucosa(6.23±0.23)(P＜0.05).There were significant differences in both of them.LVD value(38.15±0.35) in lymph metastasis group was higher than in the in the non- lymph metastasis group(27.56±0.05)(P＜0.05).There were significant differences in both of them.Conclusions:1,Expression of VEGF-D in the normal tissues was low.Expression of VEGF-D in the squamous cell carcinoma of tongue was high and no correlation with age,tumor diameter,histologic types. 2,Expression of VEGFR-3 was observed in normal tissues and tumor.But expression of VEGFR-3 in the squamous cell carcinoma of tongue was higher.Its high expression was correlation with the high expression of VEGF-D in the squamous cell carcinoma of tongue.Its high expression was also correlation with lymph node metastasis.3,The D2-40 marked positive lymphatic vessels was mainly observed in the squamous cell carcinoma of tongue and also observed in the normal tissues.But the density of lymphatic vessel of the squamous cell carcinoma of tongue was higher than the normal tissues.The density of lymphatic vessels had thin wall,irregular lumens and shapes.It mainly distributed in the tumor borderline.The density of lymphatic vessel of the squamous cell carcinoma of tongue was correlation with lymph node metastasis.4,The combined detection of VEGF-D/VEGFR-3/LVD in the squamous cell carcinoma of tongue could be accepted as important affiliated markers to evaluate the lymph node metastasis.It also could measure the prognosis of squamous cell carcinoma of tongue and provide experimental and theoretical foundation to confirm the clinlcal treatment methods.