Shatrova A N, Aksenov N D, Poletaev A I, Zenin V V
Institute of Cytology RAS, St. Petersburg.
Tsitologiia. 2003;45(1):59-68.
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) and enabled us to discriminate cells with 4c DNA--G2, M, including metaphase and anaphase cells and cells in pseudo-G1. 1 microM etoposide evoked cell accumulation in G2 phase, while 40 microM 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, and after that treated with 50 microM etoposide or 20 microM amsacrine, were shown to enter pseudo-G1 with 4c DNA amount per cell. In the presence of nocodazole, 4 and 40 microM 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 microM amsacrine in the presence of nocodazole, followed by incubation in drug-free medium, resulted in the similar transition of cells to pseudo-G1.
已表明拓扑异构酶II(拓扑II)抑制剂依托泊苷和安吖啶会导致GM - 130和Hep - 2细胞积累4c DNA量。基于流式细胞术的差异分析(Zenin等人,2001年)使我们能够区分具有4c DNA的细胞——G2期、M期细胞,包括中期和后期细胞以及假G1期细胞。1微摩尔依托泊苷引起细胞在G2期积累,而40微摩尔依托泊苷则阻断细胞增殖,这通过有丝分裂细胞和4c DNA细胞积累的完全缺失得到证实。首先用诺考达唑阻断然后冲洗的GM - 130和Hep - 2细胞,之后用50微摩尔依托泊苷或20微摩尔安吖啶处理,结果显示每个细胞以4c DNA量进入假G1期。在诺考达唑存在的情况下,4微摩尔和40微摩尔安吖啶在1小时内使所有有丝分裂细胞转变为假G1期。在诺考达唑存在下用40微摩尔安吖啶对GM - 130细胞进行15或30分钟的脉冲处理,随后在无药物培养基中孵育,导致细胞向假G1期发生类似转变。