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Expression of functionally P-glycoprotein in MA104 kidney cells
M A Capella1, M Orind, M M Morales
1Laboratório de Fisiologia Renal, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Brazil.
Zeitschrift Fur Naturforschung. C, Journal of Biosciences
|March 31, 1999
Summary
MA104 cells, a type of rhesus monkey kidney cell, exhibit unusual ouabain resistance due to high expression of P-glycoprotein (P-gp). This P-gp expression was confirmed through multiple methods, indicating its functional role in these cells.
Area of Science:
- Cell Biology
- Biochemistry
- Molecular Biology
Background:
- MA104 cells, derived from rhesus monkey kidneys, form a polarized epithelium.
- These cells display unique characteristics, including resistance to ouabain despite normal Na(+)-K(+)-ATPase drug affinity.
Purpose of the Study:
- To investigate the unusual drug resistance in MA104 cells.
- To determine the expression and function of P-glycoprotein (P-gp) in MA104 cells.
Main Methods:
- Drug resistance assays using vincristine and P-gp modulators (verapamil, cyclosporin A, trifluoperazine).
- Rhodamine 123 accumulation studies.
- Reverse transcription-polymerase chain reaction (RT-PCR) for mdr1-mRNA detection and sequencing.
- Western blot and immunofluorescence for P-gp protein expression.
Main Results:
- MA104 cells showed strong resistance to vincristine, reversible by P-gp modulators.
- Low accumulation of rhodamine 123 was observed, which increased with verapamil treatment.
- RT-PCR detected mdr1-mRNA with 96% sequence identity to human mdr1.
- Western blot and immunofluorescence confirmed P-gp expression in the cell membrane.
Conclusions:
- MA104 cells intrinsically express high levels of functional P-glycoprotein in their membranes.
- The observed drug resistance mechanisms in MA104 cells are attributed to P-gp activity.