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Journal Article
Research Support, Non-U.S. Gov't
Identification of a NF κ B Inhibition Site on the Proximal Promoter Region of Human Organic Anion Transporting Polypeptide 1A2 Coding Gene SLCO1A2 .
Organic anion transporting polypeptides (OATPs; gene symbol SLCO ) are membrane transporters that mediate the transport of wide ranges of compounds. The expression of different OATP members has been reported in the kidney, liver, placenta, brain, and intestine. Because of their broad substrate spectra and wide distribution within the human body, these transporters have been proposed to play key roles in the influx transport of many oral drugs. Inflammation is known to regulate the expression and functions of many drug-metabolizing enzymes and drug transporters. As a proinflammatory cytokine, tumor necrosis factor- α (TNF α ) has been shown to affect the expression of different drug transporters, including OATP family members. In the present study, a putative nuclear factor- κ B (NF κ B) binding site ranging from -1845 to -1836 was identified at the proximal promoter region of OATP1A2 coding gene SLCO1A2 Electrophoretic mobility shift assays and chromatin immunoprecipitation showed that nuclear extracts from both breast cancer cell MCF7 and liver cancer cell HepG2 interacted with an oligonucleotide probe containing the putative NF κ B binding site and that the DNA-protein complexes contained both p65 and p50 subunits of NF κ B. Further study revealed that the binding site may be responsible in part for the suppression effect of TNF α toward SLCO1A2 expression because the treatment of TNF α significantly increased. Treatment of TNF α significantly increased formation of the DNA-protein complexes and mutations at essential bases of the putative NF κ B binding site abolished responsiveness to the TNF α neutralizing antibody, suggesting that the binding site may be responsible in part for the suppression effect of TNF α towars SLCO1A2 expression.
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