Phosphorylation Of FBXO11 By NDR1 Inhibits Epithelial-Mesenchymal Transition In Prostate Cancer Cells

Objective:To explore the mechanism of NDR1 affecting the EMT process of prostate cancer cells and the metastasis of prostate cancer.To clarify the specific molecular mechanism of NDR1 regulating FBXO11 and β-catenin in prostate cancer cells.Methods:Through database query and analysis,we summarized the relationship between the expression levels of NDR1,FBXO11 and β-catenin and the progression and metastasis of prostate cancer;molecular cloning and plasmid transfection technology changed the expression levels and protein activity of NDR1 and FBXO11 in cells;Use cells that change the expression level of NDR1 or FBXO11 to perform cell scratch and Transwell phenotypic experiments to verify the effects of the two molecules on the migration and invasion of prostate cancer cells;RT-qPCR and Western Blot methods detect the mRNA and the corresponding molecules in the cells Protein level expression;IP and immunofluorescence experiments verify the interaction between NDR1,FBXO11,and β-catenin;protein stability,post-IP phosphorylation and ubiquitination experiments verify the phosphorylation regulation of FBXO11 and βcatenin Ubiquitination regulation mechanism;using NDR1 overexpressing and FBXO11 silenced cells to construct a mouse prostate cancer metastasis model,immunohistochemistry verified that NDR1 affects prostate cancer metastasis by regulating FBXO11.Results:1.The gene expression of NDR1 and FBXO11 decreases with the progression of prostate cancer,while β-catenin is the opposite.2.The overexpression of NDR1 and FBXO11 can not only inhibit the migration and invasion of prostate cancer cells,but also inhibit the cell EMT process.3.NDR1 can indirectly increase the ubiquitination level of β-catenin.4.FBXO11 can directly bind to β-catenin at the protein level and increase the ubiquitination of β-catenin,and this process is regulated by NDR1.5.NDR1 can directly bind to FBXO11 and phosphorylate certain serine and threonine of FBXO11.6.The phosphorylation of FBXO11 by NDR1 is necessary for NDR1 to regulate β-catenin ubiquitination.7.In animal experiments,the inhibition of prostate cancer metastasis by NDR1 can be achieved by regulating FBXO11.Conclusion:1.Both NDR1 and FBXO11 play the role of tumor suppressor genes in prostate cancer.2.The inhibition of NDR1 and FBXO11 on prostate cancer metastasis is achieved by affecting the EMT process of tumor cells.3.In prostate cancer cells,NDR1 activates FBXO11 through phosphorylation so that FBXO11 binds to βcatenin and promotes the ubiquitination and degradation of β-catenin,thereby inhibiting the EMT process of tumor cells and reducing prostate cancer metastasis.