台盼蓝拒染法、MTT法、CCK-8法在研究As_2O_3细胞毒性作用中的意义

Significance of the research of trypan blue dye exclusion,MTT and CCK-8 methods on the cytotoxicity of As_2O_3

目的筛选三氧化二砷（As203）对HL一60细胞毒性的有效剂量及作用时间的检测方法，并对其结果进行分析。方法采用台盼蓝拒染法、M1Tr法、CCK～8法测定不同浓度的As203，体外作用HL一60细胞株的细胞存活率．采用单因素方差分析其与剂量一时间依赖反应的关系。结果4．0、8．0μmol／L作用48、72h及8．0μmol／L作用24h被染细胞小于5％，其余均未观察到蓝染细胞。MTY法检测在1．0～8．0μmol／LAs20，剂量范围内培养24、48、72h（P〈0．05），各组均能够抑制HL-60细胞活性。CCK-8法检测在0．5～8．0μmol／LAs203剂量范嗣内培养24、48、72h（P〈0．05），各组均能够抑制HL-60细胞活性。HL-60细胞在不同时间、不同浓度均能出现细胞存活率明显降低，As20，的毒副作用呈现明显的剂量-时间依赖反应。结论在研究As203的细胞毒性时，用CCK-8法优于M11’法，均比台盼蓝法更为灵敏。

Objective To screen the detection methods of effective dose and the effect-time of the toxicity of As203 on the HL-60 cells, and analyze the results. Methods The HL-60 cells were put in different concentrations of As203 in vitro, and the survival fraction of the HL-60 ceils were measured by trypan blue dye exclusion, MTI＇ and CCK-8 methods. The database and statistic analyses were performed by using one-way ANOVA with dose-time-dependent re- sponse relationship. Results The blue-stained cells were less than 5% when 4.0, 8.0 μmol/L as for the 48, 72 h and 8.0 μmol/L as for the 24 h, the rest was not observed the blue-stained cells. With MTI＂ method, there was significant difference of the activity of HL-60 cells was repressed by adding As203 in the dose range of 1.0-8.0 μmol/L as for the 24, 48 and 72 h （P 〈 0.05）. With CCK-8 method, there was significant difference of the activity of HL-60 cells was repressed by adding As203 in the dose range of 0.5-8.0 μmol/L as for the 24, 48 and 72 h （P 〈 0.05）. The cell viability of HL-60 cells at different times and different concentrations were reduced significantly, while the toxicity of As203 showed obvious dose - time-dependent response. Conclusion CCK-8 in the study of cytotoxicity of As203 is superior to MTT, and both are more sensitive than trypan blue dye exclusion method.