Saltar al contenido
Merck
  • Phenotypic and transcriptional profiling in Entamoeba histolytica reveal costs to fitness and adaptive responses associated with metronidazole resistance.

Phenotypic and transcriptional profiling in Entamoeba histolytica reveal costs to fitness and adaptive responses associated with metronidazole resistance.

Frontiers in microbiology (2015-05-23)
Gil M Penuliar, Kumiko Nakada-Tsukui, Tomoyoshi Nozaki
RESUMEN

Antimicrobial chemotherapy is critical in the fight against infectious diseases caused by Entamoeba histolytica. Among the drugs available for the treatment of amebiasis, metronidazole (MTZ) is considered the drug of choice. Recently, in vitro studies have described MTZ resistance and the potential mechanisms involved. Costs to fitness and adaptive responses associated with resistance, however, have not been investigated. In this study we generated an HM-1 derived strain resistant to 12 μM MTZ (MTZR). We examined its phenotypic and transcriptional profile to determine the consequences and mRNA level changes associated with MTZ resistance. Our results indicated increased cell size and granularity, and decreased rates in cell division, adhesion, phagocytosis, cytopathogenicity, and glucose consumption. Transcriptome analysis revealed 142 differentially expressed genes in MTZR. In contrast to other MTZ resistant parasites, MTZR did not down-regulate pyruvate:ferredoxin oxidoreductase, but showed increased expression of genes for a hypothetical protein (HP1) and several iron-sulfur flavoproteins, and downregulation of genes for leucine-rich proteins. Fisher's exact test showed 24 significantly enriched GO terms in MTZR, and a 3-way comparison of modulated genes in MTZR against those of MTZR cultured without MTZ and HM-1 cultured with MTZ, showed that 88 genes were specific to MTZR. Overall, our findings suggested that MTZ resistance is associated with specific transcriptional changes and decreased parasite virulence.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Cloruro de sodio, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Sodium chloride solution, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
Sodium chloride solution, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
Cloruro de sodio, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
L-cisteína, 97%
Sigma-Aldrich
L-cisteína, from non-animal source, BioReagent, suitable for cell culture, ≥98%
SAFC
Sodium chloride solution, 5 M
Sigma-Aldrich
N,N′-Methylenebisacrylamide, powder, for molecular biology, suitable for electrophoresis, ≥99.5%
Sigma-Aldrich
Cloruro de sodio, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
L-cisteína, BioUltra, ≥98.5% (RT)
Sigma-Aldrich
Cloruro de sodio, JIS special grade, ≥99.5%
Sigma-Aldrich
Sodium chloride solution, BioUltra, for molecular biology, ~5 M in H2O
Sigma-Aldrich
Cloruro de sodio, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
E-64, protease inhibitor
Sigma-Aldrich
Cloruro de sodio, 99.999% trace metals basis
Sigma-Aldrich
N,N′-Methylenebis(acrylamide), 99%
SAFC
L-cisteína
Sigma-Aldrich
Sodium chloride solution, 5 M
Sigma-Aldrich
N,N′-Methylenebisacrylamide solution, suitable for electrophoresis, 2% in H2O
Sigma-Aldrich
Cloruro de sodio, SAJ first grade, ≥99.0%
Sigma-Aldrich
Cloruro de sodio, meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%
Sigma-Aldrich
L-cisteína, ≥97%, FG
Supelco
Cloruro de sodio, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Cloruro de sodio, BioPerformance Certified, ≥99% (titration), suitable for insect cell culture, suitable for plant cell culture
Sigma-Aldrich
N,N′-Methylenebisacrylamide, suitable for electrophoresis (after filtration or allowing insolubles to settle)
Sigma-Aldrich
Paromomycin sulfate salt, ≥98% (TLC)
Sigma-Aldrich
Cloruro de sodio, AnhydroBeads, −10 mesh, 99.999% trace metals basis
Sigma-Aldrich
Paromomycin sulfate salt, powder, BioReagent, suitable for cell culture, potency: ≥675 μg per mg
Sigma-Aldrich
Sodium chloride-35Cl, 99 atom % 35Cl
Sigma-Aldrich
L-cisteína, produced by Wacker Chemie AG, Burghausen, Germany, ≥98.0%