Study On The Gene Expression Of Vascular Endothelial Growth Factor (VEGF) In Human Esophageal Carcinoma And The Inhibitory Effect Of Antisene VEGF RNA On The Growth Of Esophageal Cancer Cell In Vivo & In Vitro

Esophageal cancer (EC) is one of the most common malignant diseases with poor prognosis,metastasis and high mortality .Although the different available multi-modalities of therapies, including radical therpy,surgical operation,chemical therpy,intervention therapy et al, have currently been used in the treatment for esophageal cancer, the prognosis of esophageal cancer has not been markedly improved.Therefore, it has an important clinic significance to study on the mechanism of the development and progression of esophageal cancer and the new strategies for the treatment of esophageal cancer with the molecular biology. With the development of the basic research on oncological fields, it was found that many tumors have the over expression in growth factors, and progression of tumors depends on angiogenesis. The vascular endothelial growth factor (VEGF) is an important factor which elicits the angiogensis in tumors. It takes an important role in the growth,differentiation,metastasis,proliferation of tumors. The growth of esophageal cancer is highly depended on angiogenesis, so VEGF is considered to have key roles in the development of esophageal cancer. By now, the systemic researches on the relationship between the expression of VEGF and histopathologic type,angiogenesis of esophageal cancer have not been reported.And there are also no reports on gene therapy against angiogeneis of esophageal cancer. This study contains three parts of work:The first,the relationships between the expression of VEGF gene in esophageal cancer and the tumor grade, tumor metastasis, angiogenesis of esophageal cancer. The second, the inhibitory effect of VEGF antisense RNA on the growth of TE-1 esophageal cancer cell line in vivo. The third, the inhibitory effect of VEGF antisense RNA on TE-1 cell in nude mice. Part Ⅰ: Study on the expression of VEGF in human esophageal cancer Objective: To study expression of VEGF in human esophageal cancer.Methods: Intratumoral microvessel density (MVD) and the protein expression of VEGF in human esophageal cancer were studied by immunohistochemistry staining. Expression of VEGF mRNA were determined in 11 human esophageal cancer and human esophageal cancer cell lines (TE-1,TE-10) by reverse transcriptase-polymerase chain reaction(RT-PCR) method.Results: 1. The expression rates of VEGF in esophageal cancer were 70.9%. The level of VEGF protein was also significantly correlated to pathologic grades,lymph node metastasis and invading depth of the tumors.There were no correlation between VEGF expression and the clinicopathologic classification,sex,age,length. 2. MVD was also significantly associted with pathologic grades,lymph node metastasis of the tumors.There were no correlation between MVD and the clinicopathologic classification,sex,age,length,vasive depth.There were positive relationship between MVD and VEGF(r=0.456,P<0.01). 3.VEGFmRNA including VEGF189,VEGF165,VEGF121was detected in human esophageal cancer cell lines (TE-1,TE-10).VEGF121 and VEGF165 were identified as the predominant specices produced in esophageal cancer cell.Conclusions: Esophageal cancer is related to higher expression of VEGF gene. There were relationship between overexpression of VEGF in esophageal cancer and grade,metastasis ,MVD of esophageal cancer .The study provides significant basis of theory and experiment for go into researching relation between esophageal cancer and VEGF gene.Part Ⅱ The inhibitory effect of VEGF antisense RNA on the growth of esophageal cancer cells in vitroObjective: To investigate the effect of antisense VEGF mRNA on mediating proliferation of TE-1 cell and its lessening effect on the angiogenesis. Methods: 1. Vector plasmid were enlarge and adopt throught DH5α and identify by limited enzyme.2.We confirmed a/the overexpression of VEGF in TE-1 cells by RT-PCR analysis,insitu hybridization and immunohistochemistry. The fragments of VEGF antisense cDNA,empty vector plasmid DNA were transfected into TE-1 cells mediated with lipofectamine re...