title: ABCB1 regulation through LRPPRC is influenced by the methylation status of the GC -100 box in its promoter.
abstract: One of the potential mechanisms of imatinib mesylate (IM) resistance in chronic myeloid leukemia (CML) is increased level of P-glycoprotein (Pgp). Pgp is an efflux pump capable of activating the multidrug resistance (MDR) phenotype. The gene encoding Pgp (ABCB1) has several binding sites in its promoter region, along with CpG islands and GC boxes, involved in its epigenetic control. In previous work, we performed a proteomic study to identify proteins involved in IM cross-resistance in acute leukemia. Among these proteins, we identified LRPPRC as a potential regulator of ABCB1 transcription via an invMED1 binding site in ABCB1. Interestingly, this invMED1 binding site overlaps with the GC -100 box. In this work, we investigated the potential role of LRPPRC in the regulation of ABCB1 transcriptional activity in CML resistance. In addition, we evaluated the potential connection between this regulation and the methylation status of the ABCB1 promoter in its GC -100 box. Our results show that LRPPRC binds prominently to the ABCB1 promoter in Lucena cells, an IM-resistant cell line. Luciferase assays showed that ABCB1 transcription is positively regulated by LRPPRC upon its knockdown. Pyrosequencing analysis showed that the ABCB1 promoter is differentially methylated at its GC -100 box in K562 cells compared with Lucena cells, and in CML patients with different response to IM. Chromatin immunoprecipitation and Pgp expression after DNA demethylation treatment showed that LRPPRC binding is affected by the methylation status of ABCB1 GC -100 box. Taken together, our findings indicate that LRPPRC is a transcription factor related to ABCB1 expression and highlight the importance of epigenetic regulation in CML resistance.
authors.affiliation: Laboratório Célula-Tronco - CEMO; INCA; Rio de Janeiro, Brazil.
authors.forename: Stephany
authors.initials: S
authors.lastname: Corrêa
authors.affiliation: Laboratório Célula-Tronco - CEMO; INCA; Rio de Janeiro, Brazil.
authors.forename: Renata
authors.initials: R
authors.lastname: Binato
authors.affiliation: Laboratório Célula-Tronco - CEMO; INCA; Rio de Janeiro, Brazil.
authors.forename: Bárbara
authors.initials: B
authors.lastname: Du Rocher
authors.affiliation: Laboratório Célula-Tronco - CEMO; INCA; Rio de Janeiro, Brazil.
authors.forename: Gerson
authors.initials: G
authors.lastname: Ferreira
authors.affiliation: Laboratório Biologia Molecular - CEMO; INCA; Rio de Janeiro, Brazil.
authors.forename: Paola
authors.initials: P
authors.lastname: Cappelletti
authors.affiliation: Programa de Carcinogênese Molecular; INCA; Rio de Janeiro, Brazil.
authors.forename: Sheila
authors.initials: S
authors.lastname: Soares-Lima
authors.affiliation: Programa de Carcinogênese Molecular; INCA; Rio de Janeiro, Brazil.
authors.forename: Luis Felipe
authors.initials: LF
authors.lastname: Pinto
authors.affiliation: Universidade do Estado do Rio de Janeiro; UERJ; Rio de Janeiro, Brazil.
authors.forename: André
authors.initials: A
authors.lastname: Mencalha
authors.affiliation: Laboratório Célula-Tronco - CEMO; INCA; Rio de Janeiro, Brazil.
authors.forename: Eliana
authors.initials: E
authors.lastname: Abdelhay
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics / Q000378:metabolism; D000328:Adult; D000368:Aged; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D001549:Benzamides / Q000494:pharmacology / Q000627:therapeutic use; D045744:Cell Line, Tumor; D019175:DNA Methylation; D000077209:Decitabine; D018432:Drug Resistance, Multiple / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D005260:Female; D006801:Humans; D000068877:Imatinib Mesylate; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000188:drug therapy / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D010879:Piperazines / Q000494:pharmacology / Q000627:therapeutic use; D011401:Promoter Regions, Genetic; D011743:Pyrimidines / Q000494:pharmacology / Q000627:therapeutic use
chemical_list: C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D000970:Antineoplastic Agents; D001549:Benzamides; C087940:LRPPRC protein, human; D009363:Neoplasm Proteins; D010879:Piperazines; D011743:Pyrimidines; D000077209:Decitabine; D000068877:Imatinib Mesylate; D001374:Azacitidine
keywords: ABCB1; LRPPRC; chronic myeloid leukemia; methylation; multidrug resistance
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