The Biological Role Of EIF5A2 In Gallbladder Carcinoma And Its Relationship With TCM Syndromes

Background: Tumor development is a multi-step process involving many genetic and epigenetic changes.There is increasing evidence that genomic alterations obtained during tumorigenesis contribute to tumor cells escaping immune surveillance.The eukaryotic translation initiation factor 5A2(EIF5A2)is a translation factor that has important functions in the initiation and extension of translation.In addition,it also involves transcription,mRNA conversion and nuclear transport.Recent studies have shown that EIF5A2 protein is involved in the development and progression of a variety of tumors,but its role in gallbladder cancer has not been reported.Therefore,this paper studied the relationship between the expression of EIF5A2 protein and different clinical features of gallbladder carcinoma,and analyzed the clinical significance of TCM syndrome differentiation,and further explored the effect of EIF5A2 on gallbladder cancer cell lines,thus laying a theoretical foundation for the treatment of gallbladder carcinoma.Objective: Immunohistochemical staining was used to detect the expression of EIF5A2 in different gallbladder carcinoma tissues,and to investigate the effect of EIF5A2 on the occurrence,development and prognosis of gallbladder carcinoma.The effect of EIF5A2 on the biological function of gallbladder cancer cell lines was also explored at the cellular level.Thereby providing a theoretical basis for clinical diagnosis,treatment and prognosis.Methods:(1)Selecting paraffin-embedded specimens of 40 cases of gallbladder cancer resection performed in Nankai Hospital of Tianjin from January 2014 to May 2017;and purchasing gallbladder cancer tissue chips from Shanghai OUTDO Biotechnology Company Ltd.(including 40 cases patient).The clinical and pathological data of the above patients were collected.The study also conducted TCM syndrome differentiation and analyzed the clinical significance of TCM syndrome differentiation.Immunohistochemical staining was performed on the above specimens,and the effect of EIF5A2 protein expression on the development of gallbladder cancer was analyzed.(2)Analysis of the relationship between clinicopathological data and prognosis in patients with gallbladder cancer,mainly using statistical methods.Univariate analysis was performed by Keplan-Meier method,and multivariate analysis was performed on the data by Cox regression model.Survival analysis was performed on patients with gallbladder carcinoma,which provided a basis for clinical diagnosis,treatment and prognosis evaluation.(3)Transfecting gallbladder carcinoma GBC-SD cell line with si-RNA small interference fragment,thereby interfering with EIF5A2 gene,and using western blot and PCR technology to detect the knockdown efficiency.(4)The gallbladder carcinoma GBC-SD cell line with knockdown of EIF5A2 expression was used as the research object.The effects of EIF5A2 gene on the migration,cell cycle,tumorigenic ability and proliferation of human gallbladder cancer cell lines were investigated by means of plate cloning,cell cycle assay,Tranwell migration,CFSE staining and other experimental techniques.Results:(1)Immunohistochemical staining of EIF5A2 protein in pathological sections showed that the expression level of EIF5A2 was related to tumor size,tumor histology type,distant metastasis,lymph node metastasis,UICC staging and differentiation(P<0.05).There was no significant difference with age,gender,Nevin staging and Dialectical of Chinese Medicine(P>0.05).(2)Univariate analysis of prognosis showed that the expression of EIF5A2,histological type,distant metastasis,Nevin stage,and degree of differentiation were closely related to the prognosis of patients(P<0.05).But there was no statistical difference between the patient’s gender,age,tumor size,UICC stage,lymph node metastasis,and TCM syndrome differentiation.Multivariate analysis of Cox regression model showed that the degree of tumor differentiation,distant metastasis,and EIF5A2 expression were independent risk factors affecting the prognosis of patients,and the difference was statistically significant(P<0.05).(3)The results of knockdown efficiency detection by Western blotting technique and PCR technique showed that the knockdown efficiency of EIF5A2-siRNA-3 transfection group was the highest compared with con-siRNA,EIF5A2-siRNA-1,EIF5A2-siRNA-2 transfection group,and the difference was statistically significant(P<0.05).(4)Cell function test results showed that compared with the con-siRNA group,EIF5A2-siRNA-3 transfection group inhibited the biological functions of proliferation,metastasis,tumorigenic ability and cell cycle of gallbladder carcinoma GBC-SD cells.Conclusion: The high expression of EIF5A2 promotes the occurrence and development of gallbladder carcinoma and has adverse effects on the prognosis of patients.The knockdown of EIF5A2 inhibits the biological function of gallbladder cancer cell lines.Therefore,this experiment provides a theoretical basis for the diagnosis,biological treatment and prognosis of gallbladder cancer.