| Objective:To investigate the expression of ANOS-1 in different gastric epithelial lesions;Meanwhile,the relationship between ANOS-1 expression and clinicopathological features and prognosis of patients with advanced gastric adenocarcinoma was analyzed,and the correlation between ANOS-1 expression and invasion and metastasis of advanced gastric adenocarcinoma was discussed.Methods:By searching GEPIA database,we analyze the difference of ANOS-1 m RNA expression between gastric adenocarcinoma and normal gastric tissues,and the relationship between ANOS-1 m RNA expression and clinical stage and survival prognosis of gastric adenocarcinoma patients with the method of bioinformatics.The correlation between ANOS-1 and N-cadherin(CDH2),VIM,SNAIL,TWIST,ZEB1 and ZEB2 in gastric adenocarcinoma tissues was analyzed by using GEPIA database.118 cases of gastric adenocarcinoma and its adjacent tissues were selected to construct tissue microarrays(TMA).ANOS-1 and E-cadherin staining are performed by immunohistochemistry(IHC);40 cases of advanced gastric adenocarcinoma with lymph node metastasis were randomly selected and ANOS-1 staining was performed for primary foci and lymph node metastasis.ANOS-1 staining was performed on 99 cases of chronic gastritis endoscopic biopsy specimens,24 cases of low grade intraepithelial neoplasia,24 cases of high grade intraepithelial neoplasia,and 24 cases of highly differentiated gastric adenocarcinoma after endoscopic submucosal dissection(ESD).Wilcoxon rank-sum test was used to analyze the expression differences between gastric adenocarcinoma and adjacent tissues,primary foci and lymph node metastasis.Kruskal-wallis H rank-sum test was used to compare the expression of ANOS-1 in gastric epithelial lesions and the expression of ANOS-1 and E-cadherin in advanced gastric adenocarcinoma tissues with different histological types,and Kruskal-Wallis rank-sum test was used for paired comparison.Chi-square test was used to analyze the relationship between ANOS-1 expression and clinicopathological features of gastric adenocarcinoma.Kaplan-Meier method for survival analysis;Independent prognostic factors were analyzed by Cox regression model.Spearman assay was used to analyze the correlation between ANOS-1 and E-cadherin expression in advanced gastric adenocarcinoma.SPSS23.0 software was used to analyze the statistical data.Results:1.GEPIA database search showed that ANOS-1 m RNA was highly expressed in gastric adenocarcinoma tissues,and the difference was statistically significant(P<0.05).ANOS-1m RNA high expression group and low expression group had significant influence on the overall survival of patients with gastric adenocarcinoma(P<0.05).Although there was no statistically significant difference in the expression of ANOS-1 m RNA among groups with different clinical stages(F=2.14,P=0.067),the overall expression level showed a trend of gradual improvement with the increase of clinical stage,and the P value was close to the test standard of statistically significant difference.2.The high expression rates of ANOS-1 in chronic superficial gastritis,chronic atrophic gastritis,low-grade intraepithelial neoplasia,high-grade intraepithelial neoplasia and well-differentiated gastric adenocarcinoma were 10.7%,16.3%,20.8%,75.0% and79.2%,respectively,and the expression levels of ANOS-1 were significantly different(P<0.001).Anos-1 expression was significantly different between low-grade intraepithelial neoplasia and high-grade intraepithelial neoplasia(P < 0.001).There were no significant difference in the expression of ANOS-1 in chronic superficial gastritis,chronic atrophic gastritis and low-grade intraepithelial neoplasia(P>0.05).There were also no significant difference in the expression of ANOS-1 between high-grade intraepithelial neoplasia and well-differentiated adenocarcinoma(P>0.05).3.ANOS-1 was highly expressed in 118 cases of gastric adenocarcinoma,which was significantly different from that in adjacent tissues(P < 0.001).ANOS-1 expression was significantly different in 99 cases of advanced gastric adenocarcinoma compared with adjacent tissues and 19 cases of early gastric adenocarcinoma compared with adjacent tissues(P<0.05).4.Among patients with advanced gastric cancer,ANOS-1 had a higher expression level in gastric adenocarcinoma tissues of TNM stage III ～ IV,invasion depth T3 ～ T4,positive lymph node metastasis and positive vascular invasion(P<0.05),but its expression level was not correlated with gender,age,degree of differentiation and tumor size(P>0.05).5.Among the 40 patients with lymph node metastasis,24 patients showed high expression in the primary foci,while 31 patients showed high expression in the corresponding lymph node metastasis.There was statistically significant difference in the expression of ANOS-1 in the primary foci and corresponding lymph node metastasis of advanced gastric adenocarcinoma(P<0.001).6.In 99 cases of advanced gastric adenocarcinoma(30 tubular adenocarcinomas,10 papillary adenocarcinomas,27 mucinous adenocarcinomas,18 signed-ring cell carcinomas,and 14 low-adhesion carcinomas),compared with tubular adenocarcinomas(the proportion of score≥4 is 26.7%),papillary adenocarcinomas(the proportion of score≥4 is 30.0%),and mucinous adenocarcinomas(the proportion of score≥4 is 33.3%),ANOS-1 expression was significantly increased in signet-ring cell carcinoma(the proportion of score≥4 is 66.7%)and low adhesion carcinoma(the proportion of score≥4 is 78.6%)(P<0.05).Compared with tubular adenocarcinoma(the proportion of score > 3 is 53.3%),papillary adenocarcinoma(the proportion of score>3 is 40.0%)and mucinous adenocarcinoma(the proportion of score>3 is 37.0%),the expression of E-cadherin was significantly decreased in signet-ring cell carcinoma(the proportion of score > 3 is 16.7%)and low adhesion carcinoma(the proportion of score>3 is 14.3%)(P<0.05).7.Spearman correlation analysis showed that ANOS-1 was negatively correlated with E-cadherin expression in advanced gastric adenocarcinoma tissues(R =-0.851,P<0.001).8.The relationship between the expression of ANOS-1 and EMT process-related N-cadherin(CDH2),VIM,SNAIL,TWIST,ZEB1 and ZEB2 at m RNA level in gastric adenocarcinoma were analyzed by GEPIA database.ANOS-1 gene expression was positively correlated with N-cadherin,VIM,SNAIL,TWIST,ZEB1 and ZEB2 at m RNA levels(P<0.001).9.Patients with high ANOS-1 expression had shorter postoperative overall survival(OS)and disease-free survival(DFS)than those with low ANOS-1 expression(P<0.01).In patients with TNM stage III～IV,invasion depth T3～T4 and negative vascular invasion,the higher the expression level of ANOS-1,the shorter OS.10.Cox univariate regression analysis showed that the factors affecting OS after radical resection in patients with advanced gastric adenocarcinoma were as follows: ANOS-1expression level(P<0.05),TNM stage(P<0.001),lymph node metastasis(P<0.01),depth of invasion(P<0.05),degree of differentiation(P<0.05),vascular invasion(P<0.001);The influencing factors of DFS were ANOS-1 expression level(P<0.01),differentiation degree(P < 0.05),TNM stage(P < 0.01),lymph node metastasis(P < 0.01),vascular invasion(P<0.001).Cox multivariate regression analysis showed that ANOS-1 expression level and TNM stage were independent prognostic factors affecting OS(P<0.05)and DFS(P<0.05)in patients with gastric adenocarcinoma.Conclusion:1.The expression level of ANOS-1 in gastric adenocarcinoma was significantly higher than that in adjacent tissues,suggesting that ANOS-1 may be involved in the occurrence of gastric adenocarcinoma.The high expression of ANOS-1 is related to the malignant transformation process of gastric mucosa from low-grade to high-grade intraepithelial neoplasia and well-differentiated adenocarcinoma,and the high expression of ANOS-1 may not be an early molecular event in the transformation process from chronic gastritis to low-grade intraepithelial neoplasia.2.The high expression of ANOS-1 is closely related to the clinical stage of advanced gastric adenocarcinoma.Patients with advanced gastric adenocarcinoma with high expression of ANOS-1 have a poor prognosis,suggesting that ANOS-1 may be involved in the progression of gastric adenocarcinoma.It can be used as a new potential molecular marker to evaluate the prognosis of patients with advanced gastric adenocarcinoma.3.The high expression of ANOS-1 was significantly correlated with invasion and metastasis of advanced gastric adenocarcinoma.The high expression of ANOS-1 was associated with decreased adhesion between tumor cells.Analysis of the correlation between the expression of ANOS-1 and EMT-related genes in GEPIA database suggested that one of the possible mechanisms for ANOS-1 to reduce the adhesion between tumor cells is the EMT process,which provides a possible direction for us to explore the molecular mechanism in the future. |
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