Genomic instability in drug-resistant human melanoma cell lines detected by Alu-I-arbitrary-primed PCR.
Destabilization of the genome seems to be an important step in the generation of drug resistance. Since malignant melanoma is extremely resistant to chemotherapy, we used human melanoma cell lines as a model to investigate the putative role of genomic instability in the appearance of drug resistance. Drug-resistant variants were obtained with MNNG, BiCNU, doxorubicin and 6-thioguanine selection of melanoma cell lines. Genomic alterations in variant cells were detected by arbitrarily primed PCR of Alu-I digested DNA (Alu-I-AP-PCR). Two differential DNA bands from 6-TG-resistant cell variants were sequenced. One is homologous to intron 25 of the neural cell adhesion molecule L1 and the second to endogenous retroviral LTR sequences. We have shown that drug-resistant melanoma cell lines accumulate genomic alterations that are efficiently detected by Alu I-AP-PCR and that drug-resistant variants show genomic instability, including variations in LTR sequences, which may be associated with the appearance of the drug resistance phenotype.
Brkic
Gordana
G
Department of Microbiology and Immunology, Faculty of Health Sciences, Ben Gurion University, Beer Sheva 84105, Israel.
Gopas
Jacob
J
Tanic
Nicola
N
Dedovic-Tanic
Nasta
N
Benharroch
Daniel
D
Finkelstein-Jaworowsky
Eve
E
Kedar
Igar
I
Dimitrijevic
Bogomir
B
D020087:Alu Elements / Q000235:genetics*; D001483:Base Sequence; D004273:DNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D004317:Doxorubicin / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D008545:Melanoma / Q000188:drug therapy* / Q000235:genetics*; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D013866:Thioguanine / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D008769:Methylnitronitrosoguanidine; D004317:Doxorubicin; D013866:Thioguanine