1. Using the incorporation of [methyl-3H]thymidine as a proliferation marker, the effects of various nucleosides and nucleotides on endothelial LLC-MK2 cells were studied. We found that ATP, ADP, AMP and adenosine in concentrations of 10 μM or higher stimulate the proliferation of these cells. 2. Inhibition of ecto-ATPase (EC 3.6.1.15), 5´-nucleotidase (EC 3.1.3.5) or alkaline phosphatase (EC 3.1.3.1) significantly diminished the stimulatory effect of ATP, indicating that the effect is primarily caused by adenosine and not by adenine nucleotides. Also, the effect depends only on extracellular nucleosides, since inhibition of nucleoside uptake by dipyridamole has no influence on proliferation. 3. Other purine nucleotides and nucleosides (ITP, GTP, inosine and guanosine) also stimulate cell proliferation, while pyrimidine nucleotides and nucleosides (CTP, UTP, cytidine and uridine) inhibit proliferation. Furthermore, the simultaneous presence of adenosine and any of the other purine nucleosides is not entirely additive in its effect on cell proliferation. At the same time any pyrimidine nucleoside, when added together with adenosine, has the same inhibitory effect as the pyrimidine nucleoside alone. 4. Apparently these proliferative effects are neither caused by any pharmacologically known P1-purinoceptor, nor are they mediated by cyclic AMP, cyclic GMP, or D-myo-inositol 1,4,5-trisphosphate as second messenger, nor by extracellular Ca2+. 5. Therefore, we conclude that various purine and pyrimidine nucleosides can influence the proliferation of LLC-MK2 cells by acting on putative purinergic and pyrimidinergic receptors not previously described.
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June 1996
Research Article|
June 01 1996
Regulation of proliferation of LLC-MK2 cells by nucleosides and nucleotides: the role of ecto-enzymes
Raf LEMMENS
;
Raf LEMMENS
*
1Limburgs Universitair Centrum, Department MBW, Biochemistry, Universitaire Campus, B-3590 Diepenbeek, Belgium
*To whom correspondence should be addressed.
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Luc VANDUFFEL
;
Luc VANDUFFEL
1Limburgs Universitair Centrum, Department MBW, Biochemistry, Universitaire Campus, B-3590 Diepenbeek, Belgium
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Henri TEUCHY
;
Henri TEUCHY
1Limburgs Universitair Centrum, Department MBW, Biochemistry, Universitaire Campus, B-3590 Diepenbeek, Belgium
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Ognjen CULIC
Ognjen CULIC
1Limburgs Universitair Centrum, Department MBW, Biochemistry, Universitaire Campus, B-3590 Diepenbeek, Belgium
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Biochem J (1996) 316 (2): 551–557.
Article history
Received:
September 13 1995
Revision Received:
January 30 1996
Accepted:
February 01 1996
Citation
Raf LEMMENS, Luc VANDUFFEL, Henri TEUCHY, Ognjen CULIC; Regulation of proliferation of LLC-MK2 cells by nucleosides and nucleotides: the role of ecto-enzymes. Biochem J 1 June 1996; 316 (2): 551–557. doi: https://doi.org/10.1042/bj3160551
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