MTT法检测肝癌细胞化疗敏感性方法学探讨

Study of MTT Colorimetric Assay as Chemosensitivity Test for Hepatic Cancer Cell

目的 探讨MTT法检测肿瘤细胞化疗敏感性的影响因素。方法 选用肝癌细胞株SMMC772 1和BEL74 0 2 ,采用正交实验设计 ,分析MTT法检测肿瘤细胞化疗敏感性的条件因素对药敏检测结果的影响。结果 肝癌细胞数与生成的甲 月赞(formasan)的光密度 (OD)值呈线性正相关。肝癌细胞数 ,药物浓度 ,药物作用时间是影响MTT法检测肿瘤抑制率的重要因素。正交设计应用于MTT法检测肿瘤细胞化疗敏感性 ,与平行设计的检测结果相关一致。结论 MTT法检测肿瘤细胞化疗敏感性 ,具有简易、快速、重复性较好等优点 ,但检测条件 (肿瘤细胞数 ,药物浓度 ,药物作用时间 )

Objective To investigate the effects of conditions on chemosensitivity test using MTT colorimetric assay.Methods The effects of conditions on chemosensitivity test for hepatoma cells with MTT colorimetric assay were surveyed. Hepatoma cell line SMMC7721 and BEL7402 and orthogonal experiment design were adapted.Results The optical density(OD) of formasan directly correlated with the number of hepatoma cells. The formasan crystal was insoluble in aqueous medium,but could be solubilized in dimethylsulfoxide(DMSO). OD value of formasan solublized in DMSO was greater at wave length of 490nm than that of 630nm,and the peak of OD value was at 530nm. There was correlation between OD 490nm and OD 630nm. The OD value was affected by the number of hepatoma cells plated,the cultivating time,the concentration of MTT and the volume of DMSO. The interaction was shared between cell number and cultivating time. The assay of chemosensitivity test of hepatoma cells was affected by the number of hepatoma cells plated,the killing time,the drug concentration of MTT and quantity of DMSO. The interaction was shared between drug concentration and killing time,also between drug concentration and number of plated cells. The relationship existed between orthogonal and parallel experiment designs of MTT assay.Conclusion MTT assay as chemosensitivity test for hepatoma cells is a simple,rapid,well reproducible,but the assays of drug sensitivity of hepatoma cells were affected by the cell number,the drug concentration,and the killing time.