Antiproliferative properties of mycophenolic acid on human retinal pigment epithelial cells in vitro.
OBJECTIVE
Mycophenolate mofetil (MMF) is an immunosuppressive agent used in the prophylaxis of graft rejection in transplantology. Its antyproliferative effects on lymphocyte, monocytes, vascular smooth muscle cells and fibroblasts are well known, but to our knowledge there are no reports on its action on the retinal epithelial (RPE) cells in vitro.


METHODS
In all experiments we used mycophenolic acid (MPC), which is the biologically active form of MME Its activity was assessed on the cultures of immortalized non-transformed cells line from a human donor (ARPE19). Cells were seeded and incubated in vitro with deferent concentrations of MPC: 0.0025 pg/ml, 0.025 pg/ml, 0.25 pg/ml, 2.5 pg/ml, 25 pg/ml and 250 microg/ml. After 24 and 72 hours of incubation, proliferative activity was assessed by 5'-bromo-2'-deoxyuridine (BrdU) incorporation into cellular DNA and the amount of cell proliferation was determined using the 3-(4.5-dimethylthiazol-2yl)-2.5-diphenyltetrazolium bromide (MTT) assay. Additionally, to determine cytotoxicity of MPC, ARPE19 cells were grown to confluence and subsequently cultured in a serum-deficient medium and then, after 24 hours of incubation with different concentrations of MPC, the MTT test was performed.


RESULTS
The BrdU assay showed the decrease of DNA synthesis activity for increasing concentrations of MPC starting with 0.025 microg/ml to 250 microg/ml. The number of RPE cells assessed with MTF test decreased after the exposition to the drug concentrations of 25 microg/ml and 250 microg/ml after 24 and 72 hours of incubation, and additionally for the concncentrations of 0.25 microg/ml and 2.5 microg/ml after 72 hours of incubation.


CONCLUSIONS
MMF influences the proliferation of immortalized ARPE19 cells without evident cytotoxic effect.
Department Ophthalmology Medical, University of Wrocław. radosk@yahoo.com
Radosław
R
Kaczmarek
Ewa
E
Zaczyńska
Marta
M
Misiuk-Hojło
D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured / Q000187:drug effects; D004247:DNA / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects*; D006801:Humans; D007124:Immunoenzyme Techniques; D009173:Mycophenolic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D055213:Retinal Pigment Epithelium / Q000187:drug effects*; D012168:Retinal Pigments / Q000378:metabolism*
D012168:Retinal Pigments; D004247:DNA; D009173:Mycophenolic Acid