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DAB2IP suppresses tumor malignancy by inhibiting GRP75-driven p53 ubiquitination in colon cancer 1 1 Introduction 1 2 Materials and methods 2 2.1 Reagents and antibodies 2 2.2 Tissue microarray and immunohistochemistry (IHC) 2 2.3 Cell lines, culture, and cell stress assay 2 2.4 Small interfering RNA, plasmid, and vector construction 2 2.5 Transfection and establishment of stable cell lines 2 2.6 Transwell, colony formation, and apoptosis assays 2 2.7 Western blot assay and co-immunoprecipitation assay 3 2.8 Quantitative real-time PCR (qPCR) 3 2.9 Ubiquitination assay 3 2.10 Mass spectrum analysis 3 2.11 Animal study 3 2.12 Public database and bioinformatics analysis 3 2.13 Statistical analysis 4 3 Results 4 3.1 DAB2IP was positively associated with an improved prognosis of colon cancer in patients expressing wild-type p53 4 3.2 DAB2IP exerted tumor-suppressive effects and increased wild-type p53 protein levels in colon cancer cells 4 3.3 DAB2IP inhibited the degradation of p53 in a ubiquitin-proteasome-dependent manner 4 3.4 DAB2IP interacted with the ubiquitin ligase-related protein GRP75 7 3.5 DAB2IP suppressed p53 ubiquitination through competitive antagonism towards GRP75 7 3.

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UND

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nonfiction