Synchronization of the cell cycle using Lovastatin
Sonia JavanMoghadam-Kamrani, Khandan Keyomarsi
Abstract
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Sonia JavanMoghadam-Kamrani, Khandan Keyomarsi
Abstract
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Synchronization by Lovastatin arrests many cell types reversibly in the G(1) phase of the cell cycle. Here we show that Lovastatin (10 microM) mediates cell cycle arrest in human breast cancer cells, MCF-7 and MDA-MB-231, where 85% of cells accumulate in the G(1) phase of the cell cycle. Addition of mevalonate (at 100X the Lovastatin concentration) releases the cells from the G(1) arrest and allows for synchronous entry into late G(1), S and G(2)/M phases of the cell cycle. The expressions of different cyclins as a marker for different phases of the cell cycle are detected by western blot analysis and indicative of synchronous transition into each of cell cycle phases following the initial G(1) arrest. Due to its level of synchrony and high yield of synchronous populations of cells, Lovastatin method of cell synchronization can be used for examining gene expression patterns in a variety of different cell lines.
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Synchronization by Lovastatin arrests many cell types reversibly in the G(1) phase of the cell cycle. Here we show that Lovastatin (10 microM) mediates cell cycle arrest in human breast cancer cells, MCF-7 and MDA-MB-231, where 85% of cells accumulate in the G(1) phase of the cell cycle. Addition of mevalonate (at 100X the Lovastatin concentration) releases the cells from the G(1) arrest and allows for synchronous entry into late G(1), S and G(2)/M phases of the cell cycle. The expressions of different cyclins as a marker for different phases of the cell cycle are detected by western blot analysis and indicative of synchronous transition into each of cell cycle phases following the initial G(1) arrest. Due to its level of synchrony and high yield of synchronous populations of cells, Lovastatin method of cell synchronization can be used for examining gene expression patterns in a variety of different cell lines.
Key concepts: Cell synchronization, Lovastatin, Cell cycle, Biology, Cell cycle checkpoint, Cell biology, Mitosis, Cell