DNA DAMAGE, CYTO-TOXIC EFFECT AND CELL-CYCLE PERTURBATION OF HOECHST-33342 ON L1210-CELLS INVITRO
DNA DAMAGE, CYTO-TOXIC EFFECT AND CELL-CYCLE PERTURBATION OF HOECHST-33342 ON L1210-CELLS INVITRO
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DOI：
10.1002/cyto.990090102
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发表时间：
1988-01-01
期刊：
影响因子：
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通讯作者：
DINCALCI, M
中科院分区：
文献类型：
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作者：
ERBA, E;UBEZIO, P;DINCALCI, M
本研究旨在评价活体染料Hoechst 33342（HO 33342）在用于获得良好DNA直方图分辨率的浓度下对L1210细胞DNA完整性、细胞生长和细胞周期时相分布的影响。通过碱洗脱法评估，HO 33342在37 ℃暴露2小时产生DNA单链断裂。DNA单链断裂是浓度依赖性的（在0.5 -5 μ g/ml范围内），并且当HO 33342（0.5-1.5 μ g/ml）与在流式细胞仪中暴露于紫外激光束照射相关时显著增加。HO 33342即使在0.5 μ g/ml的浓度下也对细胞生长产生细胞毒性作用，该浓度比获得良好DNA直方图分辨率所需的浓度小10倍。HO 33342对细胞周期G2-M期的细胞产生严重阻滞，在染色剂暴露后24 h已经明显，并持续至恢复开始后144 h。未染色细胞中不存在的新的多倍体细胞群（具有4c DNA含量）在染料暴露后24小时已经很明显。本论文中显示的数据表明，在使用HO 33342染料染色的分选细胞进行生物学、生物医学和药理学研究时应谨慎。
This study was designed to evaluate the effects of vital dye Hoechst 33342 (HO 33342), at concentrations used to obtain a good DNA histogram resolution, on DNA integrity, cell growth, and cell-cycle phase distribution of L1210 cells. HO 33342 exposure for 2 h, at 37.degree.C produced DNA single-strand breaks as assessed by the method of alkaline elution. DNA single-strand breaks were concentration dependent (in the range .5-5 .mu.g/ml) and increased significantly when HO 33342 (0.5-1.5 .mu.g/ml) was associated with exposure in a flow cytometer to U.V. laser beam illumination. HO 33342 produced a cytotoxic effect on cell growth even at the concentration of 0.5 .mu.g/ml-a concentration ten-fold smaller than those required to obtain a good DNA histogram resolution. HO 33342 produced a severe block of the cells in the G2-M phase of the cell cycle already evident 24 h after stain exposure and continuing up to 144 h after start of recovery. A new polyploid cell population (with a 4 c DNA content) not present in the unstained clels was already evident 24h after dye exposure. The data shown in the present paper would imply caution in using sorted cells stained with HO 33342 dye for biological, biomedical, and pharmacological studies.