Cell cycle specific toxicity of the Hoechst 33342 stain in untreated or irradiated murine tumor cells.
Cell cycle specific toxicity of the Hoechst 33342 stain in untreated or irradiated murine tumor cells.
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DOI：
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发表时间：
1986-07
期刊：
影响因子：
11.2
通讯作者：
D. Siemann;P. Keng
中科院分区：
文献类型：
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作者：
D. Siemann;P. Keng
Hoechst 33342 染色剂之前已与荧光激活细胞分选结合使用，将细胞群分成细胞周期的各个阶段。本实验旨在评估该染色剂对源自固体 KHT 肉瘤的细胞的毒性，这些细胞在解离前未经处理或辐射。在未经处理和受辐射的细胞中，染色剂毒性随着暴露时间的增加而增加，但在经过处理的细胞中总是更大；例如，在 10 µM 染色剂暴露 1 小时或以 5 µM 染色剂暴露 2 小时后，受辐射细胞的存活率降低 25 至 45 倍，未处理细胞的存活率降低 4 至 5 倍。为了确定这种毒性是否在细胞周期的所有阶段均等地发生，还是优先在特定阶段发生，使用离心淘析将源自未经处理或辐射的 KHT 肿瘤的细胞分成细胞周期的各个阶段。然后将每个阶段的细胞分成两部分，一半用 5 microM Hoechst 33342 染色 2 小时，另一半以相同方式处理但不染色。选择这些特定的染色暴露条件进行详细评估，因为它们提供了最佳的 DNA 直方图。在未经处理和辐射的细胞中，Hoechst 33342 的细胞毒性作用在 S 期细胞中明显强于细胞周期 G1 和 G2-M 期细胞。这种毒性，特别是其细胞周期特异性，表明在旨在确定细胞周期各个阶段治疗效果的研究中，将 Hoechst 33342 与荧光激活细胞分选结合使用可能存在严重限制。
Hoechst 33342 stain has been used previously in conjunction with fluorescence activated cell sorting to separate cell populations into the various phases of the cell cycle. The present experiments were designed to evaluate the toxicity of this stain in cells derived from solid KHT sarcomas which were untreated or irradiated prior to dissociation. In both untreated and irradiated cells, stain toxicity increased with increasing exposure times but was always greater in treated cells; for example, after stain exposures of 1 h at 10 microM or 2 h at 5 microM, survival was reduced 25- to 45-fold in irradiated cells and 4- to 5-fold in untreated cells. To determine whether this toxicity occurred equally in all phases of the cell cycle or preferentially in a particular phase, cells derived from untreated or irradiated KHT tumors were separated into the various phases of the cell cycle using centrifugal elutriation. Cells in each phase then were divided in two, one-half being stained with 5 microM Hoechst 33342 for 2 h, the other half being handled in an identical manner but not stained. These particular stain exposure conditions were chosen for detailed evaluation because they provided the best DNA histograms. In both untreated and irradiated cells the cytotoxic effects of Hoechst 33342 were found to be significantly greater in cells in the S phase than in cells in G1 and G2-M phases of the cell cycle. This toxicity, particularly its cell cycle specificity, suggests a potentially severe limitation for the use of Hoechst 33342 in combination with fluorescence activated cell sorting in studies aimed at determining treatment efficacies in various phases of the cell cycle.