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Characterization of a multidrug-resistant chronic myeloid leukemia cell line presenting multiple resistance
Nathalia Daflon-Yunes1, Flavio Eduardo Pinto-Silva, Raphael Silveira Vidal
1Laboratório de Imunologia Tumoral, Instituto de Bioquímica Médica, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Bloco H02, Sala 003, Rio de Janeiro, 21941-590, Brazil.
Abstract:
The multidrug-resistant (MDR) phenotype is multifactorial, and cell lines presenting multiple resistance mechanisms might be good models to understand the importance of the various pathways involved. The present work characterized a MDR chronic myeloid leukemia cell line, derived from K562 through a selective process using daunorubicin. This MDR cell line was shown to be resistant to vincristine, daunorubicin, and partially resistant to imatinib. It showed a slower duplication rate. Overexpression of ABCB1 and ABCC1 was observed at the protein and functional levels and the expression of CD95, a molecule related to cell death, was reduced in the MDR cell line. Conversely, no differences were observed related to the anti-apoptotic molecule Bcl-2 or p53 expression. The activation antigen CD69 was reduced in the MDR cell line and treatment with imatinib further decreased the expressed levels. Furthermore, secretion of IL-8 was diminished in the MDR cell line. When daunorubicin-selected cells were compared to another MDR cell line, Lucena 1, derived from the same parental line K562, and selected with vincristine, a different profile was observed in relation to most aspects studied. When both cell lines were silenced for ABCB1, differences in CD69 and CD95 were maintained, despite resistance reversal. These results reinforce the idea that cell lines selected in vitro may display multiple resistance strategies that may vary with the selective agent used as well as during different steps of the selection process.
Insights
Multidrug-resistant (MDR) cancer cell lines, developed using daunorubicin, show resistance to multiple drugs and altered cell death pathways. Different selection methods yield distinct MDR strategies, highlighting the complexity of drug resistance in cancer research.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- The multidrug-resistant (MDR) phenotype in cancer is complex, involving multiple resistance mechanisms.
- Cell lines with MDR phenotypes are valuable models for studying these resistance pathways.
- Understanding variations in MDR mechanisms is crucial for developing effective cancer therapies.
Purpose of the Study:
- To characterize a novel MDR chronic myeloid leukemia (CML) cell line derived from K562 using daunorubicin selection.
- To investigate the resistance profile, duplication rate, and molecular changes in the MDR CML cell line.
- To compare the characterized MDR cell line with another MDR line (Lucena 1) selected with a different agent (vincristine).
Main Methods:
- Development of an MDR CML cell line (K562) through stepwise daunorubicin selection.
- Assessment of resistance to vincristine, daunorubicin, and imatinib.
- Analysis of protein and functional expression of ABCB1 and ABCC1 transporters.
- Evaluation of apoptosis-related molecules (CD95, Bcl-2) and p53 expression.
- Measurement of activation antigen CD69 and IL-8 secretion.
- Gene silencing of ABCB1 to assess its role in observed phenotypes.
Main Results:
- The daunorubicin-selected MDR cell line exhibited resistance to vincristine, daunorubicin, and partial resistance to imatinib, with a slower duplication rate.
- Overexpression of ABCB1 and ABCC1 was confirmed at both protein and functional levels.
- Reduced expression of CD95 (cell death molecule) and CD69 (activation antigen) was observed; imatinib treatment further decreased CD69.
- IL-8 secretion was diminished in the MDR cell line.
- Comparison with a vincristine-selected MDR line (Lucena 1) revealed distinct resistance profiles.
- ABCB1 silencing reversed drug resistance but did not fully restore CD69 and CD95 levels, suggesting other contributing factors.
Conclusions:
- In vitro selected MDR cell lines can develop multiple resistance strategies that are dependent on the selective agent and selection duration.
- ABCB1 and ABCC1 overexpression, along with reduced CD95 and CD69 expression, are key features of this daunorubicin-selected MDR CML line.
- The distinct profiles between differently selected MDR lines underscore the heterogeneity of drug resistance mechanisms in cancer.
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