Down-regulation Of TRIM29 Contributes To Proliferation And Metastasis In Esophageal Carcinoma

Background and Purpose: Esophagus cancer(EC)is one of the most common malignant tumors in the world today.It has high incidence rate and mortality rate,mainly including two pathological types: esophageal squamous cell carcinoma(ESCC)and esophageal adenocarcinoma(EAC).The prognosis of esophageal cancer is poor.It is an important method to find a target for early diagnosis and treatment of esophageal cancer.TRIM29(tripartite motif containing 29)is a member of trim protein family,which can form homodimer or heterodimer in the process of nucleic acid connection.It plays a transcriptional regulatory role in the occurrence and development of tumor.At present,studies have found that the expression of TRIM29 is significantly up-regulated in a variety of tumors,such as colorectal cancer,gastric cancer and so on,but it is low expressed in some tumors,such as prostate cancer and breast cancer,suggesting that the effect of TRIM29 may be completely opposite in different tumor backgrounds.At the same time,the expression and specific role of TRIM29 in esophageal cancer are still unclear.In this project,we combined with the bioinformatics analysis and cell biology experiment of esophageal cancer clinical specimen database,elaborated the expression of TRIM29 in esophageal cancer,explored the biological mechanism of the gene involved in the occurrence and development of esophageal cancer,and further explored its specific signaling pathway and mechanism of action.Experimental Methods:(1)Using bioinformatics methods,the Cancer Genome Atlas(TCGA)database and Oncomine database of esophageal cancer were used to analyze the expression differences of TRIM29 in adjacent tissues,benign lesions and esophageal cancer;(2)The relationship between the expression of TRIM29 in TCGA and GEO esophageal cancer data sets and the clinical features of esophageal cancer,such as TNM stage,grade,differentiation,etc.,was analyzed by bioinformatics method,and the role of TRIM29 expression in the prognosis of esophageal cancer was investigated by Kaplan-Meier analysis;(3)Analyzing the correlation between the methylation level of the promoter region of TRIM29 in the TCGA esophageal cancer data set and its expression,as well as the relationship between the expression of TRIM29 and the degree of methylation in esophageal cancer cells to investigate the regulatory mechanism of TRIM29 expression;(4)KYSE-30 control cells and TRIM29 overexpression cell lines,KYSE-410,KYSE-450 control cells and TRIM29 knockout cell lines were established by lentivirus infection;(5)The effects of TRIM29 expression on the migration,invasion,colony formation and proliferation of esophageal cancer cells were investigated by cell scratch test,Transwell migration test,colony formation test and CCK-8 proliferation test;(6)Gene set enrichment analysis(GSEA)was used to study the signal pathway related to TRIM29 expression;(7)Three groups of esophageal cancer cell lines were established to investigate the molecular mechanism of TRIM29 regulating downstream signaling pathway by Western blotting.Experimental Results:(1)The expression of TRIM29 in esophageal cancer and benign lesions was significantly lower than that in adjacent / normal esophageal cancer tissues;(2)In esophageal cancer,with the increase of TNM stage and grade,the expression of TRIM29 gradually decreased,and the lower the expression of TRIM29.In addition,TRIM29 was negatively correlated with lymph node metastasis.In conclusion,the higher the degree of malignancy,the lower the expression of TRIM29;the lower the expression of TRIM29,the worse the prognosis of patients with esophageal cancer;(3)In clinical specimens of esophageal cancer,the expression of TRIM29 was negatively correlated with the methylation level of its promoter region,and TRIM29 was regulated by methylation in esophageal cancer cells;(4)Up regulation of TRIM29 can significantly inhibit the migration,invasion,colony formation and proliferation of esophageal cancer cells;(5)GSEA gene set enrichment analysis showed that the expression of TRIM29 was positively correlated with the activity of p53 signaling pathway and apoptosis pathway,but negatively correlated with EMT signaling pathway;(6)Western blotting showed that TRIM29 could inhibit the invasion and metastasis of esophageal cancer cells by regulating IL6 / STAT3 signaling pathway and ZNF750 expression.Research Conclusions: In this study,we systematically analyzed the expression level of TRIM29 in various cancer and normal tissues,esophageal cancer and adjacent tissues,different types of tumors and their stages,EC tissues and clinicopathological variables,prognostic survival analysis and methylation site survival analysis.Objective to study the expression of TRIM29 in esophageal cancer cells and its effects on the migration,invasion,colony forming ability and proliferation of esophageal cancer cells,and further explore the molecular mechanism of TRIM29 related pathways in esophageal cancer cells.It was found that TRIM29 overexpression can significantly inhibit the migration,invasion,colony formation and proliferation of esophageal cancer cells,and TRIM29 affects the physiological behavior of esophageal cancer cells through the p53 signaling pathway and EMT signaling pathway.These studies provide new ideas for the treatment of esophageal cancer and may become a new target for the treatment of esophageal cancer.