| Background:Ovarian cancer is one of the three common malignant tumors of female genital organs,among which epithelial ovarian cancer is the most common one.As it has no obvious early symptoms and lacks effective screening and diagnosis methods,more than 70% of confirmed cases have been in advanced stage and the 5 year survival rate is only about 30%,which has been a serious threat to women's health.Therefore,it is urgent to study the pathogenesis of ovarian cancer,search for early screening and diagnosis of biomarkers,and effective therapeutic targets.14-3-3 protein is a kind of small molecule acid polypeptide with a molecular weight of about 28-33 kDa,which is highly conserved in eukaryotes.So far,more than 200 ligand binding 14-3-3 protein was confirmed,through a combination of various ligands to regulate the function of 14-3-3 protein involved in the regulation of many important cellular processes,such as cell differentiation,proliferation,signal transduction,vesicular transport etc.More and more studies have confirmed that 14-3-3 beta protein is closely related to the occurrence and development of many tumors,and plays an important role in tumor growth and progression.Based on the previous research work,this experiment was designed to detect the expression of 14-3-3 beta in serous ovarian carcinoma,and to explore its effect on the biological function of serous ovarian cancer cells.Objective:The aim of this study is to detect the expression of 14-3-3 beta protein in serous ovarian adenocarcinoma tissues,analyze the relationship between 14-3-3 beta expression and clinicopathological features of ovarian serous adenocarcinoma.The expression level of 14-3-3 beta protein in ovarian cancer cells were changed by transfection of 14-3-3 beta expression and interference plasmid to observe the influence of gene on the biological behavior of ovarian cancer cells and explore biological effects of 14-3-3 beta protein in the development of ovarian cancer.Methods:1.Expression level of 14-3-3 beta in 100 cases of ovarian serous adenocarcinoma,49 cases of ovarian serous adenomas and 54 cases of normal tubal fimbria was detected by immunohistochemistry assay to analyze the relationship between 14-3-3 beta expression and clinicopathological features of ovarian serous adenocarcinoma with statistical methods.2.The expression of 14-3-3 beta was detected by real-time PCR and Western blot after transfection of 14-3-3 beta overexpression and interference plasmid into ovarian cancer cell line HO8910.3.The function of ovarian cancer cells HO8910 explored by transfected with overexpression and RNAi plasmid: MTT assay was used to detect the effect of 14-3-3 beta on the proliferation of ovarian cancer HO8910 cells;Transwell assay was to detect the effect of 14-3-3 beta on tumor cell migration and invasion ability;The effect of 14-3-3 beta on the healing ability of tumor cells was detected by scratch test;The effect of 14-3-3 beta on cell cycle and apoptosis was detected by flow cytometry.Results:1.The results of immunohistochemistry showed that the positive expression of 14-3-3 beta in ovarian serous adenocarcinoma was 45%(45/100),the positive expression in ovarian serous adenomas was 6.5%(3/49),the positive expression in normal fallopian tube fimbria was 7.4%(4/54).The positive expression rate of 14-3-3 beta in adenocarcinoma was significantly higher than that in adenoma and oviduct,and the difference had statistical significance(P < 0.005).The positive expression of 14-3-3 beta was positively correlated with the age of the patients,but was not associated with the differentiation of tumor cells,FIGO stage and lymph node metastasis.2.Compared with the blank control group and the empty plasmid group,the expression level of 14-3-3 beta in ovarian cancer HO8910 cells transfected with overexpression plasmid was significantly up-regulated,and which in ovarian cancer HO8910 cells transfected with interfering plasmid were significantly down-regulated.3.Compared with the control group,transfection of 14-3-3 beta overexpression plasmid can promote the migration and invasion of ovarian cancer HO8910 cells and promote scratch healing,inhibit the apoptosis of cancer cells,but had no obvious effect on cell proliferation and cell cycle.4.Compared with the control group,transfected 14-3-3 beta interference plasmid can inhibit the migration and invasion of ovarian cancer HO8910 cells,slow scratch healing and promote apoptosis of cancer cells,had no significant effect on the proliferation and cell cycle of cancer cells.Conclusion:1.The positive expression rate of 14-3-3 beta in serous ovarian adenocarcinoma was significantly higher than that in adenoma and tubal umbrella tissues,which may be involved in the development of serous ovarian adenocarcinoma as an oncogene,and it has potential significance for the early diagnosis and targeted therapy of ovarian cancer.2.14-3-3 beta can promote the migration and invasion of ovarian cancer HO8910 cells,inhibit the apoptosis of cancer cells,but have no obvious effect on cell proliferation and cell cycle,indicated that 14-3-3 beta may play an important role in the biological progress of ovarian cancer. |
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