| Objective: Hepatocellular carcinoma (HCC) is the sixth most common tumor worldwide but due to its poor prognosis it ranks as the third most common cause of death from cancer. It is responsible for 662,000 deaths worldwide per year. HCC accounts for more than 90% of primary liver malignancies. HCC has a distinct geographic distribution. A high incidence is described in countries in the China.The actual incidence may go up as high as 39 cases/100,000 population in China.And, 40% of the global new HCC cases diagnosed annually come from China.Although patients with early-stage disease have an excellent prognosis with a 5-year survival of more than 75–80%, 50% of HCC patients are diagnosed at an advanced stage.And there has been no effective therapy available for those with advanced disease, who are not eligible for transarterial chemoembolisation. The diagnosis of HCC is typically made by radiological liver imaging in combination with serum AFP. The sensitivity and specificity of AFP are not satisfactory at present. The PI3K/Akt/mTOR signal pathway has an important position in its regulation of apoptosis, promote angiogenesis, tumor metastasis, proliferation and other functions. Tie-2 is a tyrosine kinase receptor mainly expression in the vascular endothelial cell.It's ligand is angiopoietins, and combind to regulating vascular angiogenesis or reconstruction , and activation of Tie-2 are PI3K/Akt/mTOR signal transduction pathway factor. Soluble endothelium-specific receptor tyrosine kinase-2(sTie-2) is generated by a shedding mechanism by which the transmembrane celluar receptor is proteolytically cleaved leading to the release of the extracellular domain. Stimulators such as Phorbol 12-myristate 13-acetate(PMA) induce a decrease of Tie-2 protein from the cell surface and then a soluble form of Tie-2 comprising parts of the extracellular domain can be detected. Although the role of sTie-2 is not clear, however, found that liver cancer patients in clinical sTie-2 serum concentration and disease stage and mortality have been reported have relevance. However, the early liver cancer serum of sTie-2 has not yet reported. The aim of this study was to evaluate the diagnostic value of the serum levels of sTie-2 in HCC.Method:1 Experimental animals 75 male SPF (specific pathogen-free) SD(Sprague-Dawley) Rats.2 Construct the animal model Animals were randomlized into 4 groups. Control group (n=10): live oil solution 0.2ml/100g, intraperitoneal injection,2 times/week, 10weeks;Experimental group A(n=10): liver injury model,50% CCl4 oil solution,0.2ml/100g,intraperitoneal injection, 2 times / week.30% alcohol, 1ml/100g ,gastric irrigation,3 times / week,6 weeks;Experimental group B(n=15), liver cirrhosis model,50% CCl4 oil solution,0.2ml/100g,intraperitoneal injection, 2 times / week.30% alcohol, 1ml/100g ,gastric irrigation,3 times / week,15 weeks;Experimental group C(n=40), hepatocellular carcinoma model, 50% CCl4 oil solution , 0.2ml/100g , intraperitoneal injection, 2 times / week, 30% alcohol, 1ml/100g ,gastric irrigation,23times / week, 23 weeks.3 General observation Daily observation of animals color, diet, faeces and activities,and recording weight and water intake.4 Serum samples Tail blood,1ml,blood coagulation after 30 minutes at room temperature, centrifuged (1000 rpm) 10 minutes at 4℃, - 70℃cryopreservation. Collet Time: control group (n = 10) the first day of the second week; experimental group A (n = 10) the first day of the seventh week; experimental group B (n = 15) the first day of the sixteenth week; experimental group C (n = 40) the first day of the twenty fourth week.5 Liver tissue samples After the collection of blood,animals were sacrificed, immediately make anatomical examination, observation of the liver size, color, texture and check liver tissue. Admitted organization with a fixed 10% formaldehyde solution.6 Serum samples tested Serum sTie-2 were analyzed by sandwich enzyme-linked immunosorbent assay(sELISA). 7 Histological examination Liver tissue routinely get 10% formalin-fixed, paraffin-embedded, sliced, hematoxylin and eosin(HE) staining for histopathological examination.8 Statistical analysis All statistical analyses were done with SPSS software package, version11.0. Expression of the results of each group to mean±standard deviation express. Number of samples in each group were compared using one-way ANOVA, Differences to p<0.05 as a standard statistical significanceResults:1 The results of serum sTie-2 Control group were 62.123ng/ml (mean, SD±6.067) , liver injury group were58.836 ng/ml (mean, SD±6.921),liver cirrhosis group were 40.858 ng/ml (mean, SD±12.585) , hepatocellular carcinoma were 31.025 ng/ml (mean, SD±16.103)。2 Compared serum sTie-2 There was a significant difference compare Liver cirrhosis group, hepatocellular carcinoma group with control group,injury group.(F=16.060,P=0.000); There was no significant difference between hepatocellular carcinoma group and liver cirrhosis group(P=0.060). There was no significant difference between the control group and the liver injury group(P=0.519).Conclusion: The use of carbon tetrachloride and consumption of liquor induced SD rat model of liver cirrhosis and hepatocellular carcinoma and the success rate, in line with the requirements of this experiment to simulate the human liver cancer and cirrhosis serological studies and histology. The experimental results showed that serum level of sTie-2 is no significant difference between the model of liver cirrhosis and hepatocellular carcinoma (P = 0.060). It's shows that serum level of sTie-2 may not be the diagnostic maker for hepatocellular carcinoma. serum level of sTie2 combind with imaging may serve as a diagnostic tool for early hepatocellular carcinoma. |
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