title: Development of resistance to vincristine and doxorubicin in neuroblastoma alters malignant properties and induces additional karyotype changes: a preclinical model.
abstract: Cytotoxic drug treatment of neuroblastoma often leads to the development of drug resistance and may be associated with increased malignancy. To study the effects of long-term cytotoxic treatment on malignant properties of tumor cells, we established 2 neuroblastoma cell sublines resistant to vincristine (VCR) and doxorubicin (DOX). Both established cell lines (UKF-NB-2(r)VCR(20) and UKF-NB-2(r)DOX(100)) were highly resistant to VCR, DOX and vice-versa but retained their sensitivity to cisplatin. UKF-NB-2(r)VCR(20) and UKF-NB-2(r)DOX(100) expressed significant amounts of P-glycoprotein, while parental cells were P-glycoprotein negative. GD2 expression was upregulated, whereas NCAM expression was decreased in both resistant cells. Spectral karyotype (SKY) analysis revealed complex aberrant karyotypes in all cell lines and additional acquired karyotype changes in both resistant cells. All cell lines harbored high levels of N-myc amplification. Compared to parental cells, UKF-NB-2(r)VCR(20) and UKF-NB-2(r)DOX(100) exhibited more than 2-fold increase in clonal growth in vitro, accelerated adhesion and transendothelial penetration and higher tumorigenicity in vivo. We conclude that development of drug resistance and acquisition of certain karyotypic alterations is associated with an increase of additional malignant properties that may contribute to the poor prognosis in advanced forms of NB. The 2 novel neuroblastoma cell sublines also provide useful models for the study of drug resistance in aggressive forms of neuroblastoma.
authors.affiliation: Department of Hematology and Oncology, Center of Pediatrics, Clinic of J. W. Goethe-University, Frankfurt am Main, Germany.
authors.forename: Rouslan
authors.initials: R
authors.lastname: Kotchetkov
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Cinatl
authors.forename: Roman
authors.initials: R
authors.lastname: Blaheta
authors.forename: Jens-Uwe
authors.initials: JU
authors.lastname: Vogel
authors.forename: Jana
authors.initials: J
authors.lastname: Karaskova
authors.forename: Jeremy
authors.initials: J
authors.lastname: Squire
authors.forename: Pablo
authors.initials: P
authors.lastname: Hernáiz Driever
authors.forename: Thomas
authors.initials: T
authors.lastname: Klingebiel
authors.forename: Jindrich
authors.initials: J
authors.lastname: Cinatl
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis / Q000235:genetics; D000782:Aneuploidy; D000818:Animals; D002455:Cell Division; D002945:Cisplatin / Q000494:pharmacology; D002999:Clone Cells / Q000473:pathology; D018450:Disease Progression; D004317:Doxorubicin / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D016259:Genes, myc; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D051379:Mice; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D009368:Neoplasm Transplantation; D019006:Neural Cell Adhesion Molecules / Q000096:biosynthesis / Q000235:genetics; D009447:Neuroblastoma / Q000235:genetics / Q000473:pathology*; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D014410:Tumor Stem Cell Assay; D014750:Vincristine / Q000494:pharmacology*
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D009363:Neoplasm Proteins; D019006:Neural Cell Adhesion Molecules; D014750:Vincristine; D004317:Doxorubicin; D002945:Cisplatin