Double blockade of cell cycle progression by coptisine in vascular smooth muscle cells.
Coptisine, an isoquinoline alkaloid isolated from rhizome of Coptis japonica, inhibits proliferation of vascular smooth muscle cells (VSMCs). The aim of this study was to evaluate the action of coptisine, along with berberine (a structurally similar isoquinoline alkaloid), on progression of the cell cycle in VSMCs. Coptisine displayed antiproliferative action against VSMCs by blocking the cell cycle at G(1) and G(2)/M phases. The G(1) block was shown by inhibition of [(3)H]thymidine incorporation into VSMCs at coptisine concentrations higher than 15 microM. The mechanism underlying the G(1) arrest involved a decrease in cyclin D1 protein, although cyclin E, A, and B were not affected by coptisine treatment. The selective reduction in cyclin D1 protein was mainly attributable to accelerated proteolysis via proteasome-dependent pathway, since it was inhibited by a proteasome inhibitor, N-carbobenzoxy-L-leucinyl-L-leucinyl-L-norleucinal (MG132) and further the mRNA level of cyclin D1, protein synthesis, and mitogen-activated protein kinase (MAPK) activity remained unaltered. The mechanism underlying the G(2)/M arrest involved partial inhibition of tubulin polymerization, which was apparent at coptisine concentration of 3 microM. Berberine arrested the cell cycle at G(1) phase via a mechanism identical with coptisine, but did not cause block at G(2)/M phase. The results demonstrate that a small difference in the structure between isoquinoline alkaloids produces a big difference in activity, and that coptisine has a unique double action in arresting the cell cycle of VSMCs.
Tanabe
H
H
Laboratory of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Mizuho-ku, Japan.
Suzuki
H
H
Mizukami
H
H
Inoue
M
M
D000818:Animals; D001012:Aorta, Abdominal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D001013:Aorta, Thoracic / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D001483:Base Sequence; D001599:Berberine / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects*; D019938:Cyclin D1 / Q000378:metabolism; D056746:Cyclin G; D056766:Cyclin G1; D016213:Cyclins / Q000187:drug effects; D017931:DNA Primers; D004261:DNA Replication / Q000187:drug effects; D006868:Hydrolysis; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014404:Tubulin / Q000187:drug effects / Q000378:metabolism
C534704:Ccng1 protein, rat; D056746:Cyclin G; D056766:Cyclin G1; D016213:Cyclins; D017931:DNA Primers; D014404:Tubulin; D001599:Berberine; D019938:Cyclin D1