| Esophageal cancer is a malignant tumor in the esophageal squamous epithelial tissue, the patient are one of the most common malignant tumors people died of esophageal cancer each year about22million. The incidence of esophageal cancer is very high in our country, There are31.04million people suffering from esophageal cancer, China accounted for16.72million. According to the census of29provinces, municipalities and autonomous regions in China from1974to1978,700,000peopel died of cancer, which died of esophageal cancer account for157000(22.4%) less than gastric cancer (22.8%). The death of esophageal cancer less than gastric cancer, age of onset more than40years, but the incidence of more young in recent years has been growing Surgical operation is the preferred therapy of esophageal cancer, Radical resection of esophageal content on the lymph node dissection, but most people who have violations of fiber layer and surrounding tissue lose the opportunity of radical resection. now,five-year survival rate is only20%-30%after radical resection. Chemotherapy and radiotherapy are relatively limited for advanced esophageal cancer.Of the endogenous angiogenesis inhibitors in the body, endostatin has the broadest anti-cancer spectrum, endostatin inhibits65different tumor types. Tumor can secret a lot of vascular endothelial growth factor to promote new vascular.But angiogenesis inhibitors work only for the tumor vasculature. Studies have shown that the effect of separate application of the angiogenesis inhibitor is so limited for treatment of advanced tumors. more and more people Pay attention to the combination of chemotherapy and angiogenesis inhibitor in recent years, studies have shown that angiogenesis inhibitors mainly have the following three aspects:1inhibiting new vascular;2promoting blood vessels mature;3degradation of the blood vessels. Thus the maturity and normalization of tumor blood vessels increase the effect of the vascular transport of chemotherapy drugs, which play a strong role in the primary tumor. At the same time the blood vessels density decreases, blood flow reduction, tumor regression. Therefore, anti-angiogenic drugs in combination with chemotherapy drugs are a new strategy for solid tumors. Developed its own novel recombinant human endostatin thorough Escherichia coli express system, cancer models. A novel recombinant human endostatin (endostar), combined with NP chemotherapy studied by scholars of our country, was approved by the State Food and Drug Administration of China in2005for the treatment of non small cell lung cancer,and has achieved good results. the positive rate of VEGF in esophageal squamous cell carcinoma is23.9%-93.3%, the expression levels were significantly higher than normal tissue, and VEGF play a role in various periods of the esophageal. Pathology grading of esophageal cancer, tumor depth of invasion, lymph node metastasis and histological differentiation is highly relevant. It has been proved that Endostar combination with chemotherapy drugs has a synergistic effect.Objective:In this study, we establish esophegeal carcinoma model to investigate the anti-tumor effects and mechanism of the recombinant human endostatin combined with paclitaxel and cisplatin on esophegeal carcinoma for providing experimental evidence for further clinical application.Methods:Esophageal cancer cells frozen in liquid nitrogen, the recovery is generally fast-melting method, the vials are removed from liquid nitrogen and immediately placed in37℃water, so rapidly melted in a minute, and Cell suspension are moved into centrifuge tube with10%fetal calf serum2ml added to the centrifuge tube,centrifuging for three minutes,800r/min, discarding the supernatant after centrifuged,Eca-109esophageal cancer cell lines were subcultured. In the growth of cells in the logarithmic phase, nude mice were used to establised esophegeal carcinoma xenograft model by injecting left anterior subcutaneous2×106esophageal cancer cells, Subcutaneous tumor molded after one week with the size of soybean,40mice were randomly divided into four groups, control group (saline), endostatin group, the combination of endostatin with paclitaxel+cisplatin group (combined group), paclitaxel+cisplatin group (chemotherapy group), each group is ten. Control group was injected with saline by tail vein;the Endostar group was injected with1.5mg/kg, d1-14; the chemotherapy group was injected with cisplatin5mg/kg, paclitaxel15mg/kg;combined group doses are same to others. We observe the daily general situation in mice, including nude mice eating, drinking and activities. The length a (mm) and short diameter b (mm) of tumor were measured with a vernier calipe after the start of drug, and calculate the mouse tumor volume (V) by the following formula:V=ab2/2, depicts the tumor growth curve, calculate the tumor volume inhibition rate=(average tumor volume of control group-average tumor volume of treatment group)/the average tumor volume of control group×100%;after the end of experiment the animals were sacrificed, and calculating the inhibition rate of tumor weight; the apoptosis rate was detected by flow cytometry, the apoptosis rate in the comparison between each experimental group whether there is a significant difference; pro-apoptotic protein Bax and anti-apoptotic protein Bcl-2 determinated by immunohistochemicalResults:1The tumor weights and inhibiting rates:The tumor weight of each group is1.654±0.040,1.360±0.037,0.610±0.029,0.445±0.070.There were signifycant differences among all groups(P<0.05). Inhibiting rate of treatment groups is53.77%,71.54%,83.13%, respectively.2Changes of tumor volume:The tumor volume of control group increased gradually,and reached1247.738+12.357mm3at22nd day.Tumor volume of treated groups increases slowly compared with the tumor volume of control group. The tumor volume did not increase obviously in combinated group; the volume was63.90±35.28mm3at22nd day. Tumor volume of chemotherapy group gradually decreased after5days, and which of endostar treated group decreased10days later. It was141.72±44.92mm3and300.90±35.28mm3at22nd day. The tumor volume in combined group dropped most. There were significant differences among all groups(P<0.05).3Apoptosis of rate:Apoptosis of rate of each group is1.380±0.274%、14.110±2.044%、28.190±0.643%、39.630±2.223%,respectively.There was significant difference between the control group and treated groups(P<0.05). Apoptosis of rate in combined group was the highest, compared with endostar-treated group or chemotherapy treated group(P<0.05).4The expression of Bcl-2and Bax:The expression of Bcl-2protein in each group was65.400±3.502%、44.700±3.466%、23.30±2.751%、5.600±1.647%respectively. There was a significant difference among each group(p<0.05) The expression of Bax protein in each group was14.600±2.011%、23.000±1.490%、43.100±3.178%、56.800±3.011%respectively. There was significant difference among each group (P<O.05).Conclusions Combination of chemotherapy and Recombinant human endostatin can significantly inhibit the growth of tumor, esophageal cancer tumor volume of combined group less than the other experimental group and control group, there are significant differences; the endostar group comparing with control group have not adverse reactions; combination with the chemotherapy and Recombinant human endostatin comparing with chemotherapy group, no increase in adverse reactions.Recombinant human endostatin in combination with chemotherapy can increased expression of pro-apoptotic protein Bax and decreased the expression anti-apoptotic protein Bcl-2there are significant differences among the control group and the other experimental group; separatere combinant human endostatin promote to increase Bax expression, the anti-apoptotic protein Bcl-2expression was decreased, compared with the control group, there are significant differences.Recombinant human endostatin in combination with chemotherapy can make esophageal cancer cell apoptosis rate increased, there are significant differences, recombinant human vascular endothelial endostatin can promote esophageal cancer cell apoptosis rate compared with the control group There was significant difference. |
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