Madin Darby bovine kidney cell synchronization by lovastatin: application to bovine herpesvirus-1 gene expression.
The number of investigations involving cell proliferation has increased rapidly in the last years. One of the major difficulties in studying cell-cycle-related events is obtaining highly synchronous cell populations without metabolic imbalance. This study demonstrates that the Madin Darby bovine kidney (MDBK) cells, a commonly used cell line in veterinary research, can be effectively synchronized using lovastatin (Lov), a drug used to treat hypercholesteremia in humans. This was demonstrated by the following results: (i) Lov inhibits cell proliferation in a dose-dependent manner; (ii) Lov synchronizes MDBK cells mainly in the G1 and secondarily in the G2+M cell-cycle phases; (iii) the cytostatic effect of Lov can be specifically inhibited by addition of mevalonate (Mev) (Lov inhibits the synthesis of Mev); (iv) removal of Lov from G1-arrested cultures, followed by addition of Mev, resulted in the synchronous recovery of DNA synthesis; and (v) 5-bromo2'-deoxyuridine incorporation experiments revealed that MDBK cells synchronization by Lov can be followed for at least 3 cycles after removal of Lov and addition of Mev. Furthermore, as an application of investigations based on the availability of synchronized MDBK, we showed that bovine herpesvirus-1 gene expression is independent on the cell cycle.
Vanderplasschen
A
A
Département de Virologie-Immunologie, Faculté de Médecine Vétérinaire, Université de Liège, Belgium.
Hanon
E
E
Benarafa
C
C
Greimers
R
R
Op De Beeck
A
A
Loncar
M
M
Pastoret
P P
PP
D000818:Animals; D002417:Cattle; D002453:Cell Cycle / Q000187:drug effects*; D002460:Cell Line / Q000166:cytology / Q000187:drug effects* / Q000821:virology*; D005434:Flow Cytometry; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D007242:Herpesvirus 1, Bovine / Q000235:genetics*; D007668:Kidney / Q000166:cytology; D008148:Lovastatin / Q000494:pharmacology*; D013997:Time Factors
D008148:Lovastatin