Related Experiment Videos
Doxorubicin-resistant, MRP1-expressing U-1285 cells are sensitive to idarubicin
Kerstin Jönsson-Videsäter1, Göran Andersson, Jonas Bergh
1Department of Medicine, Huddinge University Hospital, Karolinska Institute, Karolinska Hospital, Stockholm, Sweden.
Abstract:
A doxorubicin-resistant subline (U-1285dox(900)) was derived from the human small cell lung carcinoma cell line U-1285. U-1285dox(900) was exposed to a wide range of anticancer agents to determine its resistance profile. In contrast to U-1285 cells, the resistant subline U-1285dox(900) expressed elevated MRP1 mRNA detected by reversed transcriptase-polymerase chain reaction (RT-PCR) and MRP1 protein analyzed with Western blot. Neither MDR1 mRNA nor P-glycoprotein could be detected in the parental cell line or resistant subline. U-1285dox(900) exhibited high resistance to doxorubicin, epirubicin, daunorubicin, and vincristine, an intermediate resistance to mitoxantrone, and a low resistance to etoposide. A collateral sensitivity to cytosine arabinoside, chlorodeoxyadenosine, and melphalan was observed. The resistance could be reversed by buthionine-sulphoximine and verapamil for all tested drugs. Compared with daunorubicin, resistance to idarubicin was very low, 14-fold and 2.6-fold, respectively. This was associated with a higher accumulation due to a slower transport of idarubicin out of U-1285dox(900) cells.
Insights
A new doxorubicin-resistant lung cancer cell line (U-1285dox(900)) shows elevated MRP1 expression, conferring resistance to several chemotherapy drugs. This resistance can be reversed by specific agents, offering potential therapeutic insights.
Area of Science:
- * Molecular oncology
- * Cancer cell biology
- * Drug resistance mechanisms
Background:
- * Small cell lung carcinoma (SCLC) is an aggressive cancer with limited treatment options.
- * Acquired resistance to chemotherapy is a major challenge in SCLC treatment.
- * Multidrug resistance protein 1 (MRP1) is implicated in cellular drug efflux and resistance.
Purpose of the Study:
- * To characterize the resistance profile of a novel doxorubicin-resistant SCLC subline (U-1285dox(900)).
- * To investigate the role of MRP1 in the observed drug resistance.
- * To explore potential strategies for reversing chemoresistance.
Main Methods:
- * Derivation of a doxorubicin-resistant subline (U-1285dox(900)) from the parental U-1285 SCLC cell line.
- * Quantification of MRP1 and MDR1 mRNA using reverse transcriptase-polymerase chain reaction (RT-PCR).
- * Analysis of MRP1 and P-glycoprotein expression via Western blot.
- * Assessment of drug resistance profiles and collateral sensitivity.
- * Evaluation of resistance reversal using buthionine-sulphoximine and verapamil.
Main Results:
- * U-1285dox(900) cells exhibited significantly elevated MRP1 mRNA and protein levels compared to parental cells.
- * No detectable MDR1 mRNA or P-glycoprotein was found in either cell line.
- * The resistant subline showed high resistance to doxorubicin, epirubicin, daunorubicin, and vincristine, with intermediate resistance to mitoxantrone and low resistance to etoposide.
- * Collateral sensitivity was observed for cytosine arabinoside, chlorodeoxyadenosine, and melphalan.
- * Buthionine-sulphoximine and verapamil effectively reversed resistance to all tested drugs.
- * Idarubicin accumulation was higher due to slower transport in U-1285dox(900) cells, correlating with lower resistance compared to daunorubicin.
Conclusions:
- * MRP1 overexpression is a key mechanism of doxorubicin resistance in the U-1285 SCLC subline.
- * The U-1285dox(900) model provides a valuable tool for studying MRP1-mediated drug resistance in SCLC.
- * Targeting MRP1 or using combination therapies with resistance modulators may overcome chemoresistance in SCLC.