Abstract
HeLa is one of the oldest and most commonly used cell lines in biomedical research. Owing to the ease of which they can be effectively synchronized by various methods, HeLa cells have been used extensively for studying the cell cycle. Here, we describe several protocols for synchronizing HeLa cells from different phases of the cell cycle, including G1 phase using the HMG-CoA reductase inhibitor lovastatin, S phase with a double thymidine block procedure, and G2 phase with the CDK1 inhibitor RO-3306. Cells can also be enriched in mitosis using nocodazole and mechanical shake-off. Releasing the cells from these blocks enables researchers to follow gene expression and other events through the cell cycle. We also describe several protocols, including flow cytometry, BrdU labeling, immunoblotting, and time-lapse microscopy, for validating the synchrony of the cells and monitoring the progression of the cell cycle.
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References
Skloot R (2010) The immortal life of Henrietta Lacks. Random House, New York
McLaughlin-Drubin ME, Munger K (2009) Oncogenic activities of human papillomaviruses. Virus Res 143:195–208
Chaudhry MA, Chodosh LA, McKenna WG, Muschel RJ (2002) Gene expression profiling of HeLa cells in G1 or G2 phases. Oncogene 21:1934–1942
Zhou JY, Ma WL, Liang S, Zeng Y, Shi R, Yu HL, Xiao WW, Zheng WL (2009) Analysis of microRNA expression profiles during the cell cycle in synchronized HeLa cells. BMB Rep 42:593–598
Stumpf CR, Moreno MV, Olshen AB, Taylor BS, Ruggero D (2013) The translational landscape of the mammalian cell cycle. Mol Cell 52:574–582
Chen X, Simon ES, Xiang Y, Kachman M, Andrews PC, Wang Y (2010) Quantitative proteomics analysis of cell cycle-regulated Golgi disassembly and reassembly. J Biol Chem 285:7197–7207
Wahl AF, Donaldson KL (2001) Centrifugal elutriation to obtain synchronous populations of cells. Curr Protoc Cell Biol Chapter 8:Unit 8.5
Banfalvi G (2008) Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation. Nat Protoc 3:663–673
Vassilev LT, Tovar C, Chen S, Knezevic D, Zhao X, Sun H, Heimbrook DC, Chen L (2006) Selective small-molecule inhibitor reveals critical mitotic functions of human CDK1. Proc Natl Acad Sci U S A 103:10660–10665
Terasima T, Tolmach LJ (1963) Growth and nucleic acid synthesis in synchronously dividing populations of HeLa cells. Exp Cell Res 30:344–362
Keyomarsi K, Sandoval L, Band V, Pardee AB (1991) Synchronization of tumor and normal cells from G1 to multiple cell cycles by lovastatin. Cancer Res 51:3602–3609
Mikulski SM, Viera A, Darzynkiewicz Z, Shogen K (1992) Synergism between a novel amphibian oocyte ribonuclease and lovastatin in inducing cytostatic and cytotoxic effects in human lung and pancreatic carcinoma cell lines. Br J Cancer 66:304–310
Javanmoghadam-Kamrani S, Keyomarsi K (2008) Synchronization of the cell cycle using lovastatin. Cell Cycle 7:2434–2440
Ma HT, Tsang YH, Marxer M, Poon RYC (2009) Cyclin A2-cyclin-dependent kinase 2 cooperates with the PLK1-SCFbeta-TrCP1-EMI1-anaphase-promoting complex/cyclosome axis to promote genome reduplication in the absence of mitosis. Mol Cell Biol 29:6500–6514
Sakaue-Sawano A, Kurokawa H, Morimura T, Hanyu A, Hama H, Osawa H, Kashiwagi S, Fukami K, Miyata T, Miyoshi H et al (2008) Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. Cell 132:487–498
Acknowledgements
Related works in our laboratory were supported in part by the Research Grants Council grants 662213 and T13-607/12R to R.Y.C.P.
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Ma, H.T., Poon, R.Y.C. (2017). Synchronization of HeLa Cells. In: Banfalvi, G. (eds) Cell Cycle Synchronization. Methods in Molecular Biology, vol 1524. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6603-5_12
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DOI: https://doi.org/10.1007/978-1-4939-6603-5_12
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