Sp1 regulates osteopontin expression in SW480 human colon adenocarcinoma cells
作者： Yoji Takami , Michael B. Russell , Chengjiang Gao , Zhiyong Mi , Hongtao Guo
DOI: 10.1016/J.SURG.2007.02.015
关键词: Biology 、 Promoter 、 Sp1 transcription factor 、 Endocrinology 、 Regulation of gene expression 、 Molecular biology 、 Chromatin immunoprecipitation 、 Osteopontin 、 Internal medicine 、 Metastasis Protein 、 Electrophoretic mobility shift assay 、 Nuclear protein
摘要: Background Osteopontin (OPN) mediates cancer metastasis. Mechanisms regulating OPN expression in human colorectal are unknown. Using SW480 colon adenocarcinoma cells, we hypothesized that transcription determines expression. Methods constitutively express OPN. Transient transfection and deletion analysis of promoter (full-length 2.1 kb)-luciferase constructs identified cis-regulatory regions. Gelshift chromatin immunoprecipitation (ChIP) assays the trans-regulatory nuclear protein. vitro adhesion, migration, invasion studies, siRNA was used to determine functional effect decreased protein Results A region, nt-80 nt-108, upregulated transcription. demonstrated specific binding proteins. Competition with unlabeled mutant oligonucleotides indicated nt-94 nt-104 (TGGGCTGGGC), essential for gelshift assays. Confirmatory ChIP showed corresponding be Sp1. Sp1 ablated (si-Sp1), resulting OPN-dependent by 50%, 70%, 65%, respectively. Exogenous addition si-Sp1 cells restored indices. Conclusions In conclude mediated tumor metastasis protein, OPN, regulates correlates
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