MTS法细胞毒试验的非线性关系研究

目的　探讨MTS法细胞毒试验的细胞数 OD关系以及药物剂量 -抑制率关系 ,为更好地应用MTS法提供提示。方法　采用HL 6 0和Raji细胞株。细胞数 OD关系试验 :96孔板接种不同浓度的细胞 ,加入MTS培养 1～ 5h后于 4 90nm波长测定OD ,用SPSS 11 0软件对数据作曲线估计。细胞毒试验 :96孔板每孔接种细胞 ,加不同剂量的药物培养 72h ,加入MTS培养 3h后于 4 90nm波长测定OD ,用SPSS11 0软件对数据作曲线估计。结果　 2种细胞 5个时间点的细胞数 OD关系以三次项曲线拟合度最好 ,r2 为 0 997～1 0 0 0 ,均大于线性模型的r2 0 938～ 0 993。 3种药物对 2种细胞的剂量 -抑制率以三次项曲线拟合度最好 ,r2 为0 998～ 1 0 0 0 ,优于对数剂量 -概率单位线性模型的r20 94 8～ 0 987。结论　三次项曲线对MTS法细胞毒试验的细胞数 OD关系有很高拟合度且优于线性曲线模型。通过药物剂量 -抑制率的三次项曲线拟合方程可望得到比常用的概率单位回归法更准确的IC50 。

Study of non-linearity relation of MTS cytotoxicity assay

AIM We found in experiment that the relation of cell number-OD in MTS cytotoxicity assay is non-linearity. In this paper we probe into the relations of cell number-OD and drug dose-inhibition rate. METHODS Cells HL-60 and Raji were used in this experiment. Assay on the relation of cell number-OD: Cells of different concentrations were seeded into 96 well plates. OD was read at 490 nm after incubating with MTS for 1～5 hour. The regression curves were estimated from the data with the software SPSS 11.0. Cytotoxicity assay: Cells were seeded into 96 well plate and incubated with drug of different doses for 72 hours. OD was read at 490 nm after incubating with MTS for 3 hour. The regression curves were estimated from the data with the software SPSS 11.0. RESULTS The cubic curves fit well the relations of cell number-OD of the 2 strains of cells at 5 time points and the coefficient R2 of these cubic curves is 0.997～1.000, greater than the R2 0.938～0.993 of linear model. For the relations of dose-inhibition rate, the cubic curves also have best fitness and their R2 is 0.998～1.000 greater than the R2 0.948～0.987 of linear model of logarithm dose-probit. CONCLUSION For the relations of cell number-OD in MTS cytotoxicity assay, the fitness of cubic curves are better than the linear model. A more accurate IC 50 would be obtained by the cubic curve equation than by probit regression that is in common use.