title: Oxidative stress involvement in chemically induced differentiation of K562 cells.
abstract: The erythroid differentiation of K562 cells could be achieved by exposure to several pharmacologic agents, including hemin, butyric acid (BA), and anthracycline antitumor drugs such as aclarubicin (ACLA) and doxorubicin (DOX). When used at subtoxic concentrations, these drugs induce the overexpression of erythroid genes, leading to hemoglobinization of cells. Because anthracyclines are known to generate oxidative damage, we intended to demonstrate the involvement of an oxidative stress in the chemically induced differentiation process. The addition of antioxidants to anthracycline- and BA-induced cells decreased their growth and dramatically reduced the percentage of differentiated cells at day 3. Northern blot analysis showed that antioxidants also decrease the expression of erythroid genes and related transcription factors in induced cells. Moreover, analyses of oxidative stress markers showed that treatment with BA, ACLA, and DOX lead to a decrease in reduced glutathione and antioxidant enzymes (glutathione peroxidase [GPx], glutathione reductase [GRase], CuZn superoxide dismutase [SOD], and catalase [CAT]). In addition, DOX increased thiobarbituric acid reactants (TBARs), and MnSOD activity was decreased by BA and DOX. Finally, the production of reactive oxygen species (ROS) by differentiating agents was demonstrated using the dihydroethidium probe in a microspectrofluorometric assay. Altogether, these results strongly suggest the involvement of an oxidative stress generated by BA or anthracyclines as the first step in the irreversible differentiation process. Additionally, these results underline the differences between BA, ACLA, and DOX molecular mechanisms.
authors.affiliation: Laboratoire de Biologie Moléculaire, EA2063-MéDIAN, Faculté de Pharmacie, Reims, France. benoit.chenais@univ-reims.fr
authors.forename: B
authors.initials: B
authors.lastname: Chénais
authors.forename: M
authors.initials: M
authors.lastname: Andriollo
authors.forename: P
authors.initials: P
authors.lastname: Guiraud
authors.forename: R
authors.initials: R
authors.lastname: Belhoussine
authors.forename: P
authors.initials: P
authors.lastname: Jeannesson
mesh_terms: D015250:Aclarubicin / Q000494:pharmacology; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D001798:Blood Proteins / Q000096:biosynthesis / Q000235:genetics; D020148:Butyric Acid / Q000494:pharmacology; D002374:Catalase / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D004317:Doxorubicin / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D005978:Glutathione / Q000032:analysis; D005979:Glutathione Peroxidase / Q000032:analysis; D005980:Glutathione Reductase / Q000032:analysis; D006427:Hemin / Q000494:pharmacology; D006801:Humans; D020014:K562 Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D009363:Neoplasm Proteins / Q000032:analysis; D016877:Oxidants / Q000494:pharmacology*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011794:Quercetin / Q000494:pharmacology; D017382:Reactive Oxygen Species; D013482:Superoxide Dismutase / Q000032:analysis; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D000903:Antibiotics, Antineoplastic; D000975:Antioxidants; D001798:Blood Proteins; D009363:Neoplasm Proteins; D016877:Oxidants; D017382:Reactive Oxygen Species; D017392:Thiobarbituric Acid Reactive Substances; D014157:Transcription Factors; D020148:Butyric Acid; D006427:Hemin; D015250:Aclarubicin; D004317:Doxorubicin; D011794:Quercetin; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D005980:Glutathione Reductase; D005978:Glutathione