MITOTIC PROGRESSION OF GM-130 AND Hep-2 CELLS IN THE PRESENCE OF ETOPOSIDE AND
AMSACRINE. THE FLOW CYTOMETRY ASSAY
À. N Shatrova,1, * N. D. Aksenov,1 A. I. Poletaev,2
V. V. Zenin 1
1 Institute of Cytology RAS, St. Petersburg, and the
2 Engelhardt Institute of Molecular Biology RAS, Moscow;
* e-mail: shan@linc.cytspb.rssi.ru
It has been shown that inhibitors of topoisomerase II (topo II) etoposide and amsacrine results in accumulation
of GM-130 and Hep-2 cells with 4c DNA amount. The differential analysis based on flow cytometry (Zenin et al., 2001)
enabled us to discriminate cells with 4c DNA - G2, M, including metaphase and anaphase cells and cells in
pseudo-G1. 1 μM etoposide evoked cell accumulation in G2 phase, while 40 μM etoposide blocked
cell proliferation, which was confirmed by a complete absence of both mitotic cells and 4c DNA cell accumulation.
GM-130 and Hep-2 cells that were first blocked and then washed from nocodazole, a nd after that treated with 50
μÌ etoposide or 20 μM amsacrine, were shown to enter pseudo-G1 with 4c DNA amount per cell. In the presence of
nocodazole, 4 and 40 μM amsacrine evoked transition of all mitotic cells to pseudo-G1 within 1 h.
15 or 30 minutes pulse treatments of GM-130 cells with 40 μÌ amsacrine in the presence of nocodazole, followed by
incubation in drug-free medium, resulted in the similar transition of cells to pseudo-G1.
Key words: metaphase, anaphase, pseudo-G1, flow cytometry
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