As2O3抑制恶性纤维母细胞增殖浓度效应及特异性的体外测评

目的体外检测三氧化二砷(As2O3)对人恶性纤维母细胞株MG-63(骨肉瘤细胞)及人正常纤维母细胞株MRC-5增殖的浓度效应,评价As2O3对恶性纤维母细胞毒作用的强度及特异性。方法应用噻唑蓝(MTT)法检测5 d内0.25～8μmol/L浓度的As2O3对细胞株MG-63及MRC-5增殖的抑制率。以累积正态分布函数拟合二者的浓度-抑制率曲线,比较50%抑制浓度(IC50)。以正态分布函数拟合二者抑制率的差值,推断最大差值及对应浓度的95%可信区间。结果1μmol/L浓度的As2O3对MG-63细胞抑制率第5天达58.8%,对MRC-5细胞最高仅达27.6%,8μmol/L浓度的As2O3对MRC-5细胞的抑制率达55.9%。使用第4天的浓度及抑制率分别拟合MG-63与MRC-5细胞剂量效应曲线的累积正态分布函数,决定系数(r2)值均大于0.98,IC50分别为0.968 6μmol/L和6.103 8μmol/L,其比值为6.30。拟合MG-63及MRC-5细胞增殖抑制率差值的正态分布函数,r2=0.990 7,最大抑制率差值为25.36%,95%可信区间对应As2O3的浓度为0.487 5μmol/L和1.049 4μmol/L。结论As2O3可有效抑制人恶性纤维母细胞增殖,在特定浓度区域内具有一定的特异性(安全性)。

Measurement and Evaluation in Vitro on Concentration-Effect and Specificity of As2O3 for Inhibiting Proliferation of Human Malignant Fibroblasts

Objective To examine in vitro the concentration effect of arsenic trioxide（As2O3） on the proliferation of human malignant fibroblast cell line MG-63（osteosarcoma） and human normal fibroblast cell line MRC-5,and to evaluate its intensity and the specificity on the malignant fibroblast cell cytotoxicity action.Methods The proliferation-inhibition rate of MG-63 and MRC-5 cell lines treated with As2O3（0.25-8 μmol/L） were observed within 5 d by the methy thiazoloyl tetrazolium（MTT） assay.The accumulation normal distribution function was adopted to fit their concentration-inhibition rate curves and the 50%of inhibition concentrations（IC50） were compared.To fit the differences of concentration-inhibition rates between MG-63 and MRC-5 with the normal distribution function and to deduce the maximum difference and the 95%credibility interval of corresponding concentration of As2O3.Results The maximum inhibition rate of MG-63 cells treat with 1 μmol/L As2O3 was 58.8%on 5 d and which of MRC-5 cells only reached 27.6%;the inhibition rate of MRC-5 with 8 μmol/L As2O3 reached 55.9%.To fit the accumulative normal distribution function of the dose-effect curves of MG-63 and MRC-5 cells with the concentration-inhibition rate on 4 d,the determination coefficients（r2） all were more than 0.98,IC50 was 0.968 6 and 6.103 8 μmol/L respectively,and their ratio was 6.30.To fit the normal distribution function of the differences of concentration-inhibition rates between MG-63 and MRC-5 on 4 d（r2=0.990 7）,the maximum difference of inhibition rates was 25.36%and the orresponding concentration of As2O3 in the 95%credibility interval was 0.487 5 μmol/L,1.049 4 μmol/L respectively.Conclusion As2O3 can effectively inhibit the proliferation of human malignant fibroblast cell（osteosarcoma） with certain specificity in the specific concentration interval.