| Background and TargetEsophageal carcinoma is a malignant tumor that is a great threat to human health, and China ranks top of the world in terms of incidence and death rate of esophagaeal carcinoma, while Henan province has high incidence rate in China. Esophageal carcinoma is still the main death reasons of tumor related diseases nowadays. In the medical field, as we know more and more about the unique sites of malignant tumors, and with the treatment research of the aimed genes, as long as the development of the Immunology and molecular biology, there is no doubt that they provide a new research method for the treatment of the Esophageal carcinoma, making the process of treating Esophageal carcinoma full of hope.In the past ten years, HDACs in the effect of occruence and developmen of tumors has been very hot in research, while HDAC1 is an important type of HDACs. Gernally, acetylation and deacetylation are in balance, when the blance is lost, it can induce tumorigenesis. In abnormal conditions, especially when there is high in malignant tumors, It’s possibaly relevent to in the happening, growing and transforming tumors Some researches show that HDAC1 expresses too much in the organization of Gastric, Hepatic Cancer, Rectocolonic, Non-small cell lung cancer, Breast cancer, B-NHL etc. while there are so few essays or reports about the expression that HDAC1 is in the Esophageal carcinoma and the research about happening and development of Esophageal carcinoma.In order to further research about the relationship between the expression of HDAC1 and the occurrence and development of Esophageal carcinoma, and find out the effective ways of inhabiting the gowth and metastasis of esophageal carcinoma. In this study, the immunohistochemistry were used to detect the expression of HDAC1 protein in esophageal carcinoma tissues, HDAC1 ASODN was used to silence HDAC1 gene, the immunocytochemistry were used to detect the expression of HDAC1 protein in EC9706 cell, and hyperplasia of cells were examine by MTT. This study can provide a theoretial basis for clinical treatment of esophageal carcinoma.Material and methods1.Material:Collect 49 cases of esophageal carcinoma tissues,23 cases of atypical hyperplasia tissues of adjacent carcinoma and 49 cases of normal esophageal mucous tissues.2.Methods:2.1 The immunohistochemistry(IHC) was used to detect the expression of HDAC1 protein in various tissues.2.2 Select target spot, Design and Synthesize target spot HDAC1 antisense oligonucleotides (ASODN1, ASODN2) and an unrelated sequence oligonucliotide (N-ODN). 2.3 Transfect three different concentrations(150,200 and 250μg/ml) of ASODN1 and ASODN2 into EC9706 lipidosome and act for 24,48 and 72h respectively, Meanwhile, the control group (EC9706 cells were treated with the culture medium) was setted up.2.4 The inhibition rate of cell growth in each group and each time period were detected by MTT assay.2.5 The immunocytochemistry(ICC) was used to detect the expression of HDAC1 protein in each group cells.2.6 Statistical analysis:SPSS 13.0 software was used, including chi-square test and one-way analysis of variance, test standard a=0.05..Results1. The Expression of HDAC1 in esophageal carcinoma tissuesHDAC1 show in EC9706 cells were examined by immunohistochemistry:in the carcinoma cells were Karyon cytoplasms with brown granules. Examine the level of expression of HDAC1 protein in the clinical samples from 49 cases of esophageal carcinoma tissues,23 cases of atypical hyperplasia tissues of adjacent carcinoma, and 49 normal esophageal mucosa tissuess by IHC. It was found that the The Recombinant Protein of NEDD9 is mainly located in the nucleus of esophageal carcinoma cell and the level of expression of HDAC1 protein was decreased in turn in normal esophageal mucous tissues, atypical hyperplasia tissues of adjacent hyperplasia and normal esophageal mucous tissues. The positive rates of HDAC1 protein are 63.27%(31/49),39.13%(9/23) and 20.41%(10/49) respectively, the differences among the groups are statistically significant (P<0.05). It is found that the expression of HDAC1 protein was related to neoplasm invasiveness and lymphatic metastasis (all P<0.05)2. The Expression of NEDD9 in EC9706 cellsICC was used to detect the expression of HDAC1 protein in EC9706 cells. In contrast with control group, the expression of HDAC1 protein of each transfection group is invariably lowered and the difference is of statistical significance (P<0.05) with viscosity and time dependence. The expression of HDAC 1 protein is increased along with the increase in the viscosity of transfection and the lengthening of time. The expression differences of HDAC1 protein in each group are of invariably statistical significance (P<0.05). HDAC1 show in EC9706 cells were examined by immunohistochemistry:in the carcinoma cells were Karyon cytoplasms with brown granules.3. MTT test results:MTT was to check hyperplasia of each group cells. Compared with the normal group, In HDAC1 ASODN(HDAC1 ASODN1 and HDAC1 ASODN2) transfection group, the inhibition rates of cell growth increased with the increase in the concentration of transfection and the extension of transfection time, there were statistical significances differences between the various concentrations of transfected and between the dofferent time (all P<0.05)Conclusion1. HDAC1 protein highly expressed in esophageal cancer tissues and atypical hyperplasia tissues of adjacent carcinoma, which were correlated with tumor lymph node metastasis and the depth of infiltration. It suggests that HDAC1 is involved in the growth, development of esophageal cancer.2. Cell proliferation of EC9706 cells can be inhibited by HDAC1 ASODN right EC9706 cell protein expression of HDAC 1 significatly inhibited, and this study can provide a theoretial basis for clinical treatment of esophageal carcinoma. |
PDF Full Text Request