Identification of differentially expressed mRNA transcripts in drug-resistant versus parental human melanoma cell lines.
BACKGROUND
Malignant melanoma resistance to chemotherapy remains a major limitation to treatment. Our aim was to identify genes associated with drug resistance, in order to better understand the molecular events underlying the drug-resistant phenotype.


METHODS
A human melanoma cell line and its drug-resistant variants obtained by selection with MNNG or 6-thioguanine were used. Alterations in gene expression were characterized by differential display reverse transcription-polymerase chain reaction (DDRT-PCR). Prominent mRNA fragments present in selected variants and not in the parental cells were identified and characterized by cloning and sequencing. Differential expression was confirmed by real-time RT-PCR.


RESULTS
Three functionally distinct transcriptional products were demonstrated: the chaperonin subunit TCP 1-zeta-6A (CCT6A), the hyaluronan receptor CD44 and LPPR-2, the lipid phosphate phosphatase-related protein type-2.


CONCLUSIONS
Genes with altered expression were identified in drug-resistant variants. The identified molecules may provide new insights into the molecular basis for melanoma resistance to chemotherapy.
Tanic
Nikola
N
Department of Neurobiology and Immunology, Institute of Biological Research "Sinica Stankovic", Belgrade, Israel.
Brkic
Gordana
G
Dimitrijevic
Bogomir
B
Dedovic-Tanic
Nasta
N
Gefen
Nir
N
Benharroch
Daniel
D
Gopas
Jacob
J
D001483:Base Sequence; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D019008:Drug Resistance, Neoplasm; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D008545:Melanoma / Q000188:drug therapy* / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D015203:Reproducibility of Results
D018076:DNA, Complementary; D004273:DNA, Neoplasm; D012333:RNA, Messenger