title: A new form of amphotericin B - the complex with copper (II) ions - downregulates sTNFR1 shedding and changes the activity of genes involved in TNF-induced pathways: AmB-Cu2+downregulates sTNFR1 shedding and changes the activity of genes involved in TNF-induced pathways.
abstract: BACKGROUND
  A new form of amphotericin B (AmB)- complex with copper (II) ions (AmB-Cu2+) - is less toxic to human renal cells. Cytokines, including Tumor Necrosis Factor (TNF), are responsible for nephrotoxicity observed in patients treated with AmB. Another problem during therapy is the occurrence of oxidized forms of AmB (AmB-ox) in patients' circulation. To elucidate the molecular mechanism responsible for the reduction of the toxicity of AmB-Cu2+, we evaluated the expression of genes encoding TNF and its receptors alongside encoding proteins involved in TNF-induced signalization.
  
  
  METHODS
  Renal cells (RPTECs) were treated with AmB, AmB-Cu2+or AmB-ox. The expression of TNF and its receptors was evaluated by ELISA tests and real-time RT-qPCR. The expression of TNF-related genes was appointed using oligonucleotide microarrays.
  
  
  RESULTS
  Only sTNFR1 was detected, and its level was lower in AmB-Cu2+- and AmB-ox-treated cells. TNFR1 mRNA was downregulated in AmB-ox, while TNFR2 mRNA was upregulated in AmB and AmB-Cu2+. Several changes in the expression of TNF-related genes coincided with changes in the expression of TNF receptors.
  
  
  CONCLUSIONS
  The lower toxicity of AmB-Cu2+could result from the changes in the expression of TNF receptors, which coincided with the changes in the expression of genes encoding proteins involved in TNF-induced pathways. This situation might subsequently result in a changes in intracellular signalization and influence the toxicity of tested forms of AmB on renal cells.
authors.affiliation: Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Katowice, Poland. Electronic address: jgola@sum.edu.pl.
authors.forename: Joanna
authors.initials: J
authors.lastname: Gola
authors.affiliation: Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Katowice, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Strzałka-Mrozik
authors.affiliation: Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Katowice, Poland.
authors.forename: Celina
authors.initials: C
authors.lastname: Kruszniewska-Rajs
authors.affiliation: Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Katowice, Poland.
authors.forename: Adrian
authors.initials: A
authors.lastname: Janiszewski
authors.affiliation: Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Katowice, Poland.
authors.forename: Bartłomiej
authors.initials: B
authors.lastname: Skowronek
authors.affiliation: Department of Cell Biology, Institute of Biology and Biotechnology, Maria Curie-Skłodowska University, Lublin, Poland.
authors.forename: Mariusz
authors.initials: M
authors.lastname: Gagoś
authors.affiliation: Department of Biophysics, University of Life Sciences in Lublin, Lublin, Poland.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Czernel
authors.affiliation: Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Katowice, Poland.
authors.forename: Urszula
authors.initials: U
authors.lastname: Mazurek
mesh_terms: D000666:Amphotericin B / Q000737:chemistry / Q000494:pharmacology*; D000935:Antifungal Agents / Q000494:pharmacology; D002460:Cell Line; D003300:Copper / Q000737:chemistry / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D047888:Receptors, Tumor Necrosis Factor, Type I; D014409:Tumor Necrosis Factor-alpha; D003300:Copper; D000666:Amphotericin B
keywords: Amphotericin B; Copper complexes; Oligonucleotide microarrays; Real-time RT-qPCR; TNF