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DETECTION
OF DRUG RESISTANCE-RELATED mRNA MOLECULES IN MELANOMA CELL LINES
BY DD-(DIFERENTIAL DISPLAY) RT-PCR
*Tanić
N, Brkić G, *Dedović-Tanić N, Geffen N, **Benharroch D, *Dimitrijević
B, **Gopas J.
Dept. of
Microbiology and Immunology, Faculty of Health Sciences, Ben Gurion
University, Beer-Sheva, Israel
* Laboratory for Radiobiology and Molecular Genetics, Institute
"Vinča" Belgrade, Serbia and Montenegro
**
Dept. of Oncology and Pathology, Soroka Medical Center, Beer Sheva,
Israel
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ABSTRACT
The
incidence of malignant melanoma is rapidly increasing in western
populations. This malignancy is well known for its high intrinsic
and acquired resistance to chemotherapy, severely limiting the efficiency
of treatment. Although several mechanisms have been described to
account for drug resistance, there is still a need to detect new
molecules involved in this process which can be targeted by novel
therapy. In our work we have selected drug resistant human melanoma
cell lines and searched for differentialy expressed RNA populations
between the parental and the drug resistant cells. Two sets of drug
resistant cells were developed from the GA melanoma cell line, one
set resistant to MNNG and the other to 6-thioguanine (6-TG). The
DD-(Differential Display) RT-PCR technique was used to detect variant
bands between the cells. Several cDNA bands were isolated, cloned
and sequenced. The following genes were identified using the GenBank
database: the 6A subunit of the TCP-1 chaperonine, CD44, semaphorin
and the ribosomal protein L3. DD RT-PCR is a cost-effective method
to detect variant gene expression. We are in the process of validating
our results at the protein level and determining their significance
in determining or maintaining the drug resistant phenotype. |
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