| Background and Objective:Colon cancer(CC)is a late-found malignant tumor of digestive system with high malignancy and poor prognosis.However,the occurrence and development mechanism of colon cancer is still unclear,and it is urgent to find effective tumor-related biomarkers and new therapeutic targets.Recent studies have shown that Glutathione S-transferase omega 2(GSTO2)is highly expressed in a variety of malignant tumors and plays an important role in tumor development and treatment.This study aims to elucidate the effect and specific mechanism of GSTO2 on the biological behavior of colon cancer cells through cytological,zoological and histological studies,so as to provide experience and ideas for understanding the pathogenesis and situation of colon cancer and explore new therapeutic targets for colon cancer.Method:The m RNA expression level and prognosis of GSTO2 in colon cancer data sets in TCGA and GEO databases were analyzed by bioinformatics.The differentially expressed genes of GSTO2 in the data set were obtained and analyzed for functional enrichment and immune infiltration.IHC and Western blotting were used to verify the protein expression level of GSTO2 in colon cancer tissues,and q RT-PCR and Western blotting were used to detect the expression level of GSTO2 in different colon cancer cell lines.The optimal interference sequence was selected to construct lentivirus plasmid by transfection of interference fragments and verification.Next,colon cancer cell lines with stable down-regulation of GSTO2 were constructed,and the changes in the proliferation ability of colon cancer cells were investigated by CCK8 and Ed U experiments.Next,the changes of cell migration ability were studied by cell scratch assay and transwell assay.Finally,a subcutaneous tumor formation model of nude mice was constructed using colon cancer cells with stable down-regulation of GSTO2.The results of in vitro experiments were verified by comparing the tumor size of nude mice in sh GSTO2#1 group and sh GSTO2 NC group.After changing the expression level of GSTO2,the activity changes of PI3K/AKT/m TOR signaling pathway related proteins in colon cancer cells were analyzed.Result:Bioinformatics analysis showed that GSTO2 was highly expressed in colon cancer tissues and correlated with poor prognosis of patients.GO and KEGG enrichment analysis showed that the high expression of GSTO2 was related to the interaction of ECM receptors and played an important role in the immune microenvironment,and may promote the progression of colon cancer through the PI3K-Akt signaling pathway.IHC,WB and other experiments showed that GSTO2 expression in colon cancer tissue specimens was significantly higher than that in paracancer tissues(p < 0.001).q RT-PCR and WB experiments showed that GSTO2 expression was the highest in HCT116 and HT29 colon cancer cell lines.After interfering GSTO2 expression in HCT116 and HT29,CCK8 and Ed U experiments showed that its proliferative activity was weakened,while cell scratch and transwell experiments showed that its migration ability was weakened.Tumor formation in nude mice further verified that the tumor size decreased significantly after interference with GSTO2 expression level.western blot analysis was performed to detect the changes of PI3K/AKT/m TOR pathologically related proteins in sh NC group and sh GSTO2#1 group.The results showed that the protein expressions of P-Mtor,p-PI3 K and P-Akt were down-regulated after the interference of GSTO2 expression in HCT116 and HT29 cells(p < 0.01).Conclusion:1.Bioinformatics analysis and clinical sample verification have shown that GSTO2 is highly expressed in colon cancer tissues and is associated with poor prognosis of patients,which may become a prognostic indicator for colon cancer patients.2.GSTO2 is a cancer-promoting factor in colon cancer.In vivo and in vitro experiments have shown that interference with GSTO2 expression inhibits the proliferation of colon cancer cells.In vitro experiments confirmed that GSTO2 can regulate the migration ability of colon cancer cells.3.GSTO2 can regulate the proliferation and migration of colon cancer cells through PI3K/AKT/m TOR pathway. |
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