Role Of DKK2 Expression In The Progression Of Bladder Cancer

BACKGROUND: Bladder cancer is the most common malignant tumor of urinary system in China.Its incidence and mortality are the first in urinary tumors.The biological mechanism of bladder cancer development is extremely complicated,and the specific mechanism of its occurrence is still unclear.Studies have confirmed that genomic genetic information,epigenetic abnormal changes,and external effects have led to the development of bladder cancer.To clarify the molecular biological mechanism of bladder cancer,to find bladder cancer-specific non-invasive molecular markers,and to find effective therapeutic targets for bladder cancer,it is of great significance to prevent and block the occurrence and progression of bladder cancer.Studies have shown that DKK2 can promote a variety of tumorigenesis,but the role in bladder cancer is unclear.The DKK protein family contains two cysteine-rich regions that participate in embryonic development through interaction with the Wnt signaling pathway.DKK2 has a two-way regulation of Wnt/?-catenin signaling pathway.This study verified the expression of DKK2 in bladder cancer patients and a series of in vitro tests.DKK2 has a certain influence on the development of bladder cancer,which may become bladder cancer.Forecast factor.METHODS: TCGA database and GEO database were used to screen differentially expressed genes in bladder cancer,select candidate genes,construct co-expression network maps of mRNA based on prognosis-related genes,and analyze the biological processes involved in these genes by GO enrichment;in 40 cases of bladder cancer tissues The differential expression of DKK2 was detected by semi-quantitative immunohistochemistry.The correlation between the expression of DKK2 and the clinicopathological parameters of bladder cancer patients was analyzed by chi-square test.The normal immortalized bladder epithelial cell line(SV-HUC)was evaluated by QRT-PCR.-1)and DKK2 expression in four bladder cancer EJ,HTB-9(5637),T24,253-JBV cell lines,and detection of differential expression of DKK2 in bladder cancer tissues and adjacent normal tissues;cell immunofluorescence detection of DKK2 expression Positioning and differential expression levels in different cell lines.In vitro experiments: Construction of DKK2 plasmid transfection promoted the expression of DKK2 in bladder cancer,CCK-8 assay to detect the effect of DKK2 on the proliferation of bladder cancer cells,cell scratch test and Transwell invasion assay were used to detect the effect of DKK2 on migration and invasion of bladder cancer cells.To clarify the role of DKK2 in the progression of bladder cancer and to explore the relevant molecular mechanisms.RESULTS: GO enrichment analysis screened TCGA database and GEO database to screen differentially expressed genes in bladder cancer,and gene candidate for DKK2 was predicted,and the biological function of DKK2 was predicted.Real-time quantitative RT-PCR(Q-PCR)was used to display the phase.Compared with normal bladder epithelial cells,DKK2 is highly expressed in bladder cancer cells and is higher in normal tissues than in normal tissues.Immunofluorescence experiments prove that DKK2 is localized in cell membrane and cytoplasm in bladder normal and cancer cell lines,and is high in cancer cells.Expression;clinical pathology correlation by semi-quantitative analysis of IHC showed that DKK2 was significantly associated with tumor size(P<0.0001),high grade(P = 0.0127),and T stage(P=0.0354),but with patient age,gender,smoking history,tumor The number and the like were irrelevant;in predicting tumor invasion(T2-T4 tumor),the area under the ROC curve(AUC)of DKK2 was 0.730.CCK-8 experiments confirmed that the abnormal expression of DKK2 affects the proliferation of bladder cancer cells.Cell scratch assay confirmed that abnormally expressed DKK2 regulates the migration ability of bladder cancer cells.Transwell invasion assay confirmed that DKK2 regulates the invasion behavior of bladder cancer cells.Conclusion: The results indicate that DKK2 is highly expressed during the development of bladder cancer,and DKK2 promotes proliferation and invasion of bladder cancer cells.Therefore,DKK2 can be a predictor of bladder cancer and provide a potential therapeutic target.