| BackgroundEsophageal cancer is one of the most common malignant tumors with poor prognosis,poor quality of life and low survival rate.In recent years,China is on the rise.At present,the commonly used treatment methods for esophageal cancer include surgery,radiotherapy and chemotherapy,comprehensive treatment and so on.In recent years,with the gradual improvement of the treatment level and the gradual improvement of the treatment concept,the quality of life of patients has improved.However,it has to be regretted that esophageal cancer is still a disease that can not achieve the ideal treatment effect.In order to explore new treatment methods,medical experts at home and abroad have never stopped the research on esophageal cancer.A large number of gene pathways and molecular mechanisms related to the pathogenesis of esophageal cancer are gradually increasing.This is one of the ways to find new treatment methods for esophageal cancer.It is generally recognized that in the process of tumor cell formation and development,on the basis of mitosis,tumor cell proliferation is carried out by monoclone or polyclonal.Many studies have proved that Cyclin B1(Ccnb1)is a regulator of cell mitosis,and it has a high level of expression in many cancers.Ccnb1 is closely related to the replication and invasion of tumor cells,that is,the cells with higher expression of ccnb1 have stronger replication and invasion ability.Circular RNA-Cyclin B1(Circ-ccnb1)is an obvious tumor suppressor.In some tumors,it can inhibit the transfer of ccnb1 into the nucleus,that is,inhibit its nuclear translocation,and limit the development of tumors.However,the relationship between the expression level of Circ-ccnb1 and clinicopathological factors related to Esophageal squamous cell carcinoma(ESCC)is not clear.Although it is known that Circ-ccnb1 can inhibit many kinds of tumors,there is no relevant report in esophageal cancer.Objective1.To compare the expression of circ-ccnb1 in adjacent normal tissues and ESCC tissues,and its relationship with gender,age,degree of differentiation,lymph node metastasis and clinical stage.2.To observe the nuclear translocation of ccnb1 after overexpression of Circ-ccnb1 in EC109 and EC9706 cells.3.To observe the effect of overexpression of Circ-ccnb1 on the proliferation,migration and invasion of esophageal squamous cell carcinoma cells EC109 and EC9706.Material1.Tissue source:the pathological specimens of 80 patients with esophageal squamous cell carcinoma and normal esophageal mucosa adjacent to carcinoma(3-5cm from tumor tissue)were collected from January 2015 to November2016 in the Second Affiliated Hospital of Zhengzhou University.2.Cell lines:normal esophageal epithelial cells(het1-a)and esophageal squamous cell carcinoma cells(EC109,EC9706).Cell source: laboratory,School of basic medicine,Zhengzhou University.Methods1.Tissue experiment: the expression of ccnb1 was detected by immunohistochemistry in 80 cases of adjacent normal esophageal mucosa and esophageal squamous cell carcinoma,and the expression of Circ-ccnb1 was detected by PCR,and the correlation with clinicopathological factors(gender,age,degree of differentiation,lymph node metastasis and clinical stage)was analyzed.2.Cell experiment: the expression of Circ-ccnb1 in normal esophageal epithelial cells(Het1-A)and esophageal squamous cell carcinoma cells(EC109,EC9706)was detected by PCR.EC109 and EC9706 cells were transfected with Circ-ccnb1 plasmid.The cells were divided into negetive control group(NC group)and transfection group(Circ-ccnb1 group),western blot(WB)was used to detect the change of the amount of ccnb1 protein transferred to the nucleus;cell proliferation test(CCK-8 method),scratch test and transwell method were used to detect the changes of cell proliferation rate,migration rate and invasion rate in the negetive control group and the transfection group.ResultsThe expression of Ccnb1 protein in normal tissues adjacent to esophageal carcinoma was lower than that in esophageal squamous cell carcinoma(P<0.05),the expression of Circ-ccnb1 in normal tissues adjacent to esophageal carcinoma was higher than that in esophageal squamous cell carcinoma tissues(P<0.05).The expression level of Circ-ccnb1 in esophageal squamous cell carcinoma was not significantly correlated with the gender,age and degree of differentiation of the patients(P>0.05),but had a certain correlation with lymph node metastasis and clinical stage(P<0.05).The expression level of Circ-ccnb1 in Het1-A cells was higher than that in EC109 and EC9706 cells(P<0.05).Compared with the negative control group,the nuclear translocation level of Ccnb1 protein in Circ-ccnb1 overexpression group was significantly reduced(P<0.05),the proliferation rate,migration rate and invasion rate of cells were all reduced(P<0.05).ConclusionCcnb1 was highly expressed in esophageal squamous cell carcinoma,but lowly expressed in adjacent tissues.Circ-ccnb1 was highly expressed in normal esophageal adjacent tissues,but lowly expressed in esophageal carcinoma tissues.The content of Circ-ccnb1 in patients without lymph node metastasis was higher than that in patients with lymph node metastasis;the content of Circ-ccnb1 in patients with low clinical stage was higher than that in patients with high clinical stage.The content of Circ-ccnb1 in normal cells was higher than that in ESCC cells.Circ-ccnb1 can effectively inhibit the nuclear translocation of Ccnb1 in ESCC cells.Overexpression of Circ-ccnb1 can inhibit the proliferation,migration and invasion of ESCC.1.The expression of Circ-ccnb1 was high in normal esophageal cancer tissues,but low in esophageal cancer tissues.The Circ-ccnb1 level in patients without lymph node metastasis was higher than that in patients with lymph node metastasis;The level of Circ-ccnb1 in patients with low clinical stage was higher than that in patients with high clinical stage.2.Circ-ccnb1 can effectively inhibit the nuclear translocation of Ccnb1 in EC109 and EC9706 cells.3.Overexpression of Circ-ccnb1 in Ec109 and EC9706 can inhibit the proliferation,migration and invasion of ESCC. |
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