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Development of Resistance to Endoplasmic Reticulum Stress-Inducing Agents in Mouse Leukemic L1210 Cells
Martin Cagala1, Lucia Pavlikova1, Mario Seres1
1Institute of Molecular Physiology and Genetics, Centre of Biosciences, Slovak Academy of Sciences, Dúbravská cesta 9, 84505 Bratislava, Slovakia.
Abstract:
Four new variants of L1210 cells resistant to endoplasmic reticulum (ER) stressors, tunicamycin (STun), thapsigargin (SThap), bortezomib (SBor), and MG-132 (SMG-132), were developed via an 18-month periodic cultivation in culture medium with a gradual increase in substance concentration. Multidrug resistance was generated for STun (to tunicamycin, bortezomib and MG-132), SThap (to tunicamycin, thapsigargin and MG-132), SBor (to bortezomib and MG-132), and SMG-132 (to bortezomib and MG-132). These cells were compared to the original L1210 cells and another two variants, which expressed P-gp due to induction with vincristine or transfection with the gene encoding P-gp, in terms of the following properties: sensitivity to either vincristine or the ER stressors listed above, proliferative activity, expression of resistance markers and proteins involved in the ER stress response, and proteasome activity. The resistance of the new cell variants to ER stressors was accompanied by a decreased proliferation rate and increased proteasome activity. The most consistent change in protein expression was the elevation of GRP78/BiP at the mRNA and protein levels in all resistant variants of L1210 cells. In conclusion, the mechanisms of resistance to these stressors have certain common features, but there are also specific differences.
Insights
New L1210 cell variants resistant to endoplasmic reticulum (ER) stressors were developed. Resistance correlated with reduced proliferation and elevated GRP78/BiP, suggesting common and distinct resistance mechanisms.
Area of Science:
- Cell Biology
- Molecular Biology
- Drug Resistance
Background:
- Endoplasmic reticulum (ER) stress is implicated in various cellular processes and diseases.
- Developing cell models resistant to ER stressors is crucial for understanding cellular defense mechanisms.
- Multidrug resistance (MDR) is a significant challenge in cancer therapy.
Purpose of the Study:
- To generate and characterize L1210 cell variants resistant to ER stressors (tunicamycin, thapsigargin, bortezomib, MG-132).
- To investigate the cross-resistance patterns and underlying molecular mechanisms.
- To compare these novel variants with existing P-gp expressing cells.
Main Methods:
- Long-term cultivation of L1210 cells with gradually increasing concentrations of ER stressors.
- Assessment of sensitivity to various drugs and ER stressors.
- Analysis of proliferative activity, resistance markers, and ER stress response proteins.
- Measurement of proteasome activity.
Main Results:
- Four resistant L1210 cell variants (STun, SThap, SBor, SMG-132) were successfully developed.
- Acquired resistance to ER stressors was associated with decreased proliferation and increased proteasome activity.
- Elevated GRP78/BiP expression at both mRNA and protein levels was observed in all resistant variants.
- Cross-resistance patterns varied among the developed cell lines.
Conclusions:
- Resistance to ER stressors in L1210 cells involves common and specific molecular mechanisms.
- GRP78/BiP upregulation is a consistent feature of ER stress resistance in these models.
- These resistant cell lines serve as valuable tools for studying ER stress response and MDR.
