http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#Head http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c http://www.nanopub.org/nschema#hasAssertion http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#assertion http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c http://www.nanopub.org/nschema#hasProvenance http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#provenance http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c http://www.nanopub.org/nschema#hasPublicationInfo http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#pubinfo http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c http://www.w3.org/1999/02/22-rdf-syntax-ns#type http://www.nanopub.org/nschema#Nanopublication http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#assertion http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_1 http://purl.obolibrary.org/obo/RO_0002204 http://www.genenames.org/cgi-bin/gene_symbol_report?hgnc_id=10452 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_1 http://www.w3.org/1999/02/22-rdf-syntax-ns#type http://www.ebi.ac.uk/chebi/searchId.do?chebiId=CHEBI_33697 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_2 http://purl.obolibrary.org/obo/BFO_0000066 http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=9606 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_2 http://www.w3.org/1999/02/22-rdf-syntax-ns#object http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_1 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_2 http://www.w3.org/1999/02/22-rdf-syntax-ns#predicate http://www.selventa.com/vocabulary/increases http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_2 http://www.w3.org/1999/02/22-rdf-syntax-ns#subject http://www.ebi.ac.uk/chebi/searchId.do?chebiId=5295 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_2 http://www.w3.org/1999/02/22-rdf-syntax-ns#type http://www.w3.org/1999/02/22-rdf-syntax-ns#Statement http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#assertion http://www.w3.org/2000/01/rdf-schema#label a(CHEBI:gemcitabine) -> r(HGNC:RRM2) http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#provenance http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://purl.org/dc/elements/1.1/description Approximately 61,000 statements. http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://purl.org/dc/elements/1.1/rights Copyright (c) 2011-2012, Selventa. All rights reserved. http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://purl.org/dc/elements/1.1/title BEL Framework Large Corpus Document http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://purl.org/pav/authoredBy http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_4 http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://purl.org/pav/version 1.4 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_3 http://www.w3.org/ns/prov#value In our previous publication it was shown that a Gemcitabine-resistant KBGem clone derived from step-wise exposure to Gemcitabine resulted in overexpression of the human Ribonucleotide Reductase M2 subunit (hRRM2) mRNA and protein (Goan et al., 1999). In this study we confirm these results and show that the hRRM2 gene amplification arises in a homogeneous staining region (hsr) derived from chromosome translocation. The hydroxyurea-resistant clone (KBHURs) was studied as a comparison. PCR analysis of the hRRM2 gene promoter confirmed the amplification. Northern and Western blots were further employed to confirm the gene amplification and hRRM2 mRNA and protein expression were compatible with the level of drug resistance. Cells synchronized by serum starvation and then returned to serum-containing growth conditions showed a rapid induction of high levels of transcription of the hRRM2 gene. To clarify whether expression of hRRM2 mRNA was regulated at a transcriptional level, several transcription factors, including AP-1, Sp1, AP-2, CREB, NF-kappa B, and OCT1, were examined by gel-shift assay. Interestingly, the KBGem clone was regulated by different transcription factors than the KBHURs clone. Compared to the wild-type KB cells (KBwt), the KBGem clone exhibited a different binding pattern for Sp1 and NF-kappa B. The KBHURs clone, however, demonstrated a unique binding pattern with AP-1 and CREB, different from the KBwt control as well as the KBGem clone. Therefore, we conclude that the drug-resistant phenotype is associated with human RRM2 gene amplification from a homogeneous staining chromosome region and altered transcription regulation. Each clone demonstrated a unique pattern of transcription factor binding that may play a vital role in the mechanism of drug resistance. http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_3 http://www.w3.org/ns/prov#wasQuotedFrom http://www.ncbi.nlm.nih.gov/pubmed/11978967 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_4 http://www.w3.org/2000/01/rdf-schema#label Selventa http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#assertion http://www.w3.org/ns/prov#hadPrimarySource http://www.ncbi.nlm.nih.gov/pubmed/11978967 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#_3 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c#pubinfo http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c http://purl.org/dc/terms/created 2014-07-03T14:30:02.756+02:00 http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c http://purl.org/pav/createdBy http://orcid.org/0000-0001-6818-334X http://www.tkuhn.ch/bel2nanopub/RAofcycmyRA9mHAOBSE6mThM2dBxZMpO7LRY5Ze5oR40c http://purl.org/pav/createdBy http://orcid.org/0000-0002-1267-0234