ZNF326: A Potential Prognostic Biomarker For Gastric Cancer And Inhibits Cell Migration,invasion And Growth

Objective: Gastric cancer(GC),as one of the most common malignant tumors,seriously threatened human health worldwide.Although surgical treatment and other adjuvant treatments continue to improve,the prognosis of GC is still frustrating.Therefore,exploring the complex molecular mechanisms in the occurrence and development of GC,discovering efficient molecular markers for early diagnosis and prognostic evaluation of GC,and finding sites for targeted therapy have important clinical significance.The transcription initiation process of eukaryotes is very complicated and often requires the assistance of variety protein factors.Transcription factors are a group of protein molecules that can specifically participate in the process of transcription initiation.Zinc finger protein is a type of transcription factor with finger-like domains,which plays an important role in gene regulation.Many studies have found that the expression levels of multiple zinc finger proteins are important for the prognosis of patients with GC clinical significance.This reveals the mechanism of zinc finger protein in the occurrence and development of GC will help to find new biomarkers for GC.This study aims to find prognostic biomarkers for gastric cancer from the public database,verify their expression and function,select appropriate prognostic genes for gastric cancer in future clinical practice.Materials and methods: In the first part,we used GEO,TCGA databases and local sequencing data to analyze the differential expression of GC genes and their role in GC patient prognosis.According to the results of sequencing and microarray data,ZNF326 was selected as our main research object,and its tumor mutation burden,microsatellite instability,and stem cell index were analyzed in the TCGA database.Then,we analyze the expression level of ZNF326 in GC tissue by immunohistochemical staining,and analyze its correlation with patient pathological and prognostic information,verify and explore its value and significance for clinical diagnosis.In the second part,the effects of ZNF326 on the proliferation,migration and invasion of GC cells were measured by CCK8 cell proliferation test,Transwell cell migration,invasion test and wound healing test.Through nude mouse subcutaneous tumor formation and lung metastasis model experiments,the effect of ZNF326 on the function of GC cells in vivo was verified.The effect of ZNF326 on the EMT phenotype of GC cells was studied by western blotting.Results: 1.According to public databases,ZNF326 is significantly negatively correlated with the prognosis of GC patients whether in m RNA expression level or protein expression level,p <0.001,that is,those with higher relative expression in cancer have better prognosis and are independent The prognostic factor in the TNM stage;2.In the TCGA data set,the expression of ZNF326 was positively correlated with TMB(p<0.001,R=0.17),and positively correlated with MSI(p<0.001,R=0.28),which was related to the expression data based on The calculated stem cell index is positively correlated(p<0.001,R=0.23),but has no significant correlation with the stem cell index based on methylation data;3.ZNF326 is significantly low expressed in gastric cancer tissue(P=0.008),but it has nothing to do with the patient’s clinicopathological data;4.ZNF326 is a prognostic factor of GC independent of TNM stage;5.In vivo and in vitro functional experiments,overexpression of ZNF326 can inhibit the proliferation,migration and invasion of GC cells,while knockout promotes GC cells Proliferation,migration and invasion;5.ZNF326 can affect the EMT phenotype of GC cells.Overexpression of ZNF326 can inhibit the transformation of GC cells to mesenchymal type.On the contrary,knocking out ZNF326 can lead to the loss of epithelial traits of GC cells.Conclusions: 1.ZNF326 is an independent prognostic factor in GC in the public database;2.ZNF326 is significantly low in GC tissues and is a prognostic factor independent of TNM stage;3.ZNF326 inhibits tumor in migration and invasion phenotypes in both vivo and vitro,it induces GC proliferation,migration and invasion;4.ZNF326 is related to the EMT phenotype.