A Primary Study For The Association Between Lymphangiogenesis And Lymph Node Metastasis In The Esophageal Carcinoma

Background and objectiveLymph node metastasis has significance to prognosis of EC. There is some query on lymphangiogenesis and lymph node metastasis. This study was designed to investigate the expression of VEGF-C, VEGF-D,VEGFR-3, D2-40 in 91 cases of esophageal carcinoma and 20 cases of fresh specimens. The research may provide some basement of rationale to lymphangiogenesis and lymph node metastasis.Materials and methods1. Tissue samples:(1) Storaged paraffin specimens:91 cases of esophageal carcinoma and 14 cases of margin of EC paraffin specimens were collected who underwent esophageal cancer from the department of pathology of Shantou university medical college from November 2001 to November 2006.(2) Fresh specimens:20 cases of ESCC were collected from operation. Specimens were all fixed in formaldehyde immediately.2. HE stain:All the paraffin specimens were to make tissue morph and pathological diagnosis after HE staining.3. Immunhistochemistry and analysis: The Envision Labeled Peroxidase System two steps method were performed in the study. The protein expression of VEGF-C, VEGFR-3,VEGF-D, D2-40 were tested. Cells were counted by Leica Qwin standard Y2.8 Image Processing and Analysis System.4. In situ hybridization: Expression of VEGF-D mRNA was examined and its relationship with VEGF-D protein was tested by consistency checking.Result 1. Histopathology:There was no significant difference between lymph node metastasis with different years of age, gender, invasion depth(Pï¹¥0.05). The positive rate of lymph node metastasis in small cell carcinoma was significantly higher in ESCC than small cell carcinoma(P <0.05).2 .ImmunhistochemistryVEGF-C,VEGF-D,VEGFR-3,D2-40 expressed same in kinds of type of esophageal carcinoma. There was difference in lymph node metastasis with VEGF-C,VEGF-D,D2-40,VEGFR-3(P<0.05),The amount of D2-40,VEGFR-3 was higher in ESCC than normal esophageal mucosa(P<0.01). The amount of vessel VEGFR-3 expression was higher in positive group of VEGF-C,VEGF-D than negative group(P<0.05). D2-40 was higher in VEGF-C positive group,but not in VEGF-D positive group. There was difference in the amount of VEGFR-3 and D2-40.3 .In situ hybridizationThe expression of VEGF-D protein using immunhistochemistry was concordance with VEGF-D mRNA using in situ hybridization in 20 samples of ESCC(Kappa=0.737,P<0.01).4 .Image analysis:The rate of VEGF-D positive cell was higher in lymph node metastasis group than without lymph node metastasis(P<0.01).Conclusion1. VEGF-C promote Lymphangiogenesis facilitate lymph node metastasis through receptor VEGFR-3,combine with LVD,may be select as markers to prognose lymph node metastasis.2.VEGF-D was secreted by tumor cell,through receptor VEGFR-3 facilitate lymph node metastasis.3. The LVD was higher in ESCC than margin,positive group of lymph node metastasis was higher than negative group,ESCC may have lymphangiogenesis.4. VEGFR-3 expressed not only in lymphatic vessel but also in blood vessel. There was difference compare with D2-40,D2-40 may be more reliable marker of lymphatic vessel.