Synchronization of Tumor and Normal Cells from G1 to Multiple Cell Cycles by Lovastatin

Khandan Keyomarsi, Larue Sandoval, Vimla Band, Arthur B. Pardee

Research output: Contribution to journalArticle

411 Citations (Scopus)

Abstract

Synchronization of mammalian cells is essential for investigations involving cell proliferation. A simple method for obtaining synchrony in all types of cells, through several cycles and with minimal overall metabolic perturbations, has not yet been available. We describe a procedure for synchronizing normal as well as tumor cells reversibly in the GI phase of the cell cycle using Lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A reducãase. This method of synchronization was successful with all cell lines tested, including normal and tumor cells of mouse, hamster, and human origins. For example, when MCF-7 human breast cancer cells were synchronized with Lovastatin and released by the addition of mevalonic acid (the product of the reaction catalyzed by 3-hydroxy-3-methylglutaryl-coenzyme A reducãase), 3 phases of accel erated ih\rumino incorporation into DNA corresponding to 3 S phases of the cell cycle occurred during a 90-h period of cell replication. Thymidine incorporation was decreased to â€-¢4'; during the initial lag of 18 h before the first S phase, and maximum incorporation was then achieved after only 6 h. The antibody Ki-67, which detects a nuclear antigen associated with proliferation, was present in cells arrested with Lovastatin. This fact, together with the lack of thymidine incorporation during the initial lag time, indicates that the cells were arrested in the d and not in the Co phase of the cell cycle. Furthermore, in synchronized tumor-derived human breast epithelial cells, histone H4 RNA was low after Lovastatin release and increased with the onset of DNA synthesis. Concomitant synthesis of DNA and histone H4 RNA expression could be observed for 2 cycles. Minimal perturbations of general metabolic functions oc curred since the rate of RNA, protein, and initial DNA synthesis were unaffected by Lovastatin, as evidenced by |'H|uridine, ['HJIeiidne, and initial

Original languageEnglish (US)
Pages (from-to)3602-3609
Number of pages8
JournalCancer Research
Volume51
Issue number13
StatePublished - Jan 1 1991

Fingerprint

Lovastatin
Cell Cycle
Neoplasms
DNA
RNA
S Phase
Histones
Thymidine
Mevalonic Acid
Nuclear Antigens
Uridine
Cricetinae
Breast
Epithelial Cells
Cell Proliferation
Breast Neoplasms
Cell Line
Antibodies

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Keyomarsi, K., Sandoval, L., Band, V., & Pardee, A. B. (1991). Synchronization of Tumor and Normal Cells from G1 to Multiple Cell Cycles by Lovastatin. Cancer Research, 51(13), 3602-3609.

Synchronization of Tumor and Normal Cells from G1 to Multiple Cell Cycles by Lovastatin. / Keyomarsi, Khandan; Sandoval, Larue; Band, Vimla; Pardee, Arthur B.

In: Cancer Research, Vol. 51, No. 13, 01.01.1991, p. 3602-3609.

Research output: Contribution to journalArticle

Keyomarsi, K, Sandoval, L, Band, V & Pardee, AB 1991, 'Synchronization of Tumor and Normal Cells from G1 to Multiple Cell Cycles by Lovastatin', Cancer Research, vol. 51, no. 13, pp. 3602-3609.
Keyomarsi, Khandan ; Sandoval, Larue ; Band, Vimla ; Pardee, Arthur B. / Synchronization of Tumor and Normal Cells from G1 to Multiple Cell Cycles by Lovastatin. In: Cancer Research. 1991 ; Vol. 51, No. 13. pp. 3602-3609.
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