Expression Of EphA1/ephrinA3 In Esophageal Carcinoma And Their Relationship With Clinical Parameters

World wide, high incidence areas of esophageal cancer include:China, South America, Western Europe, South Africa, Japan and the former Soviet Union. From the 90's, due to the efficient intervention to the risk factors of esophageal cancer, the incidence and mortality of this cancer have been decreased gradually in Henan Province, still higher than the average level of the whole country. Eph gene is the secretion of erythropoietin in human hepatocellular carcinoma cell lines with high expression to be named, belongs to receptor tyrosine kinase gene family in the largest subgroup, involved in cell growth, differentiation, embryo development and intracellular signaling the process of delivery. They have very important physiological functions. EphAl is the first member of the receptor tyrosine kinase family, the gene is located on chromosome 7q34. ephrinA3 is EphAl one of the major ligands, has been found in human cancer its expression level is variable, but EphAl and ephrinA3 study in esophageal cancer is rare.ObjectivesBy analysis the relationship between the expression of ephrinA3, EphA1 mRNA and protien and patiens pathological parameters, to find out the two molecules of clinical significance. In esophageal cancer tissues the expression of EphAl and ephrinA3 were detected on the protein level, using esophageal cancer cell lines, study preliminarily the cell biology functions of ephrinA3 in esophageal cancer, investigate the function and clinical significance of ephrinA3 and EphAl at different levels and views.Materials and methods1 Subjects and samplesSubjects were hospitalized patients diagosed as esophageal cancer waiting for surgery in Linxian Tumor Hospital from 2007 to 2008. The patients did not receiving any forms of chemotherapy and radiotherapy treatments. Total 92 samples were collected. The gerneral information of the subjects were collected. The clinical information was also collected. All of the data were for statistically analyzed.2 Expression of EphA1/ephrinA3 mRNA in samplesReal time-PCR was used to determine the mRNA expression levels of ephrinA3/EphAl in the esophageal cancer tissues and matched normal esophageal tissues.3 EphAl and ephrinA3 protein levels in samples and ephrinA3 determination of cell growth in esophageal cancerBy western blot and immunohistochemical determination of EphAl and ephrinA3 protein expression levels in esophageal carcinoma and its matched adjacent normal tissue. In 8 human esophageal cancer cell line protein expression was detected in ephrinA3 level, and finally the selected ephrinA3 the lowest expression level of the end of TE-12 cell line, build ephrinA3 stable expression strains. Then, by using MTT assay and cell growth curve, detection ephrinA3 on esophageal cancer cell growth.4 Date analysisSPSS 12.0 statistical analysis software was used to analyze the data, t-test and correlation analysis was used for△Ct value. Theχ2-test were adopted to study the relationship between the ephrinA3/EphAl gene mRNA and protein expression and the clinical pathology factors. a=0.05 as the test standard.Results1 Real time-PCR results1.1 Expression of EphAl/ephrinA3 mRNA in esophageal cancer tissues and matched normal esophagus tissuesReal-time PCR showed that compared with matched normal esophageal tissues, in 92 patients with esophageal cancer, the expression of EphAl mRNA were down-regulated in 82(89.13%) of esophageal cancer tissues compared with the paired normal esophageal tissues, the expression of ephrinA3 mRNA were down-regulated in 53 (57.61%) of esophageal cancer tissues. Paired-Samples t-test showed that the difference of expression of EphAl mRNA have statistical significance between cancer tissues and the paired normal ones (t=-6.899, P<0.01), ephrinA3 mRNA expression in esophageal carcinoma and matched normal esophageal tissues statistically significant differences (t=-2.436, P<0.05)1.2 Relationship between expression level of ephrinA3/EphAl mRNA in esophageal cancer and the individual clinical and pathological characteristicsExpression of EphAl in the patients with family history was significantly lower than those without family history(χ2=5.838, P<0.05). The difference of EphA1 expression at different pathogical type was significant (χ2=9.660, P<0.01) Expression of ephrinA3 in the patients with family history was significantly lower than those without family history (χ2=5.417,P<0.05), Expression of ephrinA3 in the patients with matastasis was significantly lower than those without matastasis (χ2=20.504, P<0.01), The difference of ephrinA3 expression at different pathogical type was significant (χ2=6.434, P<0.05)2 Western blot, immuonhistochemistry results and relationship between protein expression of ephrinA3/EphA1 in esophageal cancer and the individual clinical and pathological characteristicsExpression of EphA1 in the patients with family history was significantly lower than those without family histor (χ2=11.482, P<0.01). The difference of EphA1 expression at different pathogical type was significant (χ2=12.818, P<0.01) Expression of ephrinA3 in the patients with family history was significantly lower than those without family history (χ2=10.391, P<0.01), Expression of ephrinA3 in the patients with matastasis was significantly lower than those without matastasis (χ2=13.615, P<0.01), The difference of ephrinA3 expression at different pathogical type was significant (χ2=7.525, P<0.05)3 ephrinA3 expression in human esophageal cancer cell lines Using Western blot method, we also analyzed the ephrinA3 expression in eight stains of human esophageal cancer cell lines. The results showed that:TE-12, Ec-9706, KYSE-180 had lower levels of expression compared with others. We choose TE-12 which had strong ability of tumor formation overexpressed ephrinA3, to further study its biological functions.Conclusion1 ephrinA3 and EphAl mRNA and protein expression levels in esophageal cancer organizations were significantly lower than esophageal normal organizations. The expression levels of ephrinA3 and EphAl mRNA were significantly negative correlated, and both maybe interaction in the process of tumorigenesis and development.2 ephrinA3 mRNA and protein expression levels closely related family history, Matastasis, Pathogical type. EphAl mRNA and protein expression levels closely related to family history, pathogical type.3 In cell experiments suggests ephrinA3 gene can inhibition esophageal cancer cell proliferation.4 ephrinA3 and EphAl genes can be used as judge the related gene transfer esophageal patients, Therefore ephrinA3 might be provide the potential treatment for the esophageal.