Kubota T, Takahara T, Nagata M, Furukawa T, Kase S, Tanino H, Ishibiki K, Kitajima M
Department of Surgery, School of Medicine, Keio University, Tokyo, Japan.
J Surg Oncol. 1993 Feb;52(2):83-8. doi: 10.1002/jso.2930520205.
A colorimetric chemosensitivity test was investigated using sulforhodamine B (SRB), which stains protein synthesized by cells, as an end-point marker. Four cultured cell lines, 9 human tumor xenografts serially transplanted into nude mice, and 14 fresh surgical specimens were subjected to this assay. The optimal conditions for the assay were 3-5 x 10(4) cells per well in a 96-microplate, an SRB concentration of 4%, and an incubation time of more than 10 minutes. When mitomycin C, doxorubicin, cisplatin, and 5-fluorouracil were assessed by the SRB assay, the concentration-effect curves revealed a sharp slope between plateaux at low and high concentrations, suggesting that this assay has an excellent sensitivity which can assess the effect of drugs as "all or none." Although this high sensitivity resulted in good reproducibility of the assay for cultured cell lines, the predictive rate of the SRB assay for the chemosensitivity of human tumor xenografts in vivo was limited to 63.9%. As a result, this SRB assay is thought to be useful for evaluating the chemosensitivity of cultured cells as all or none, since it can assess directly cellular protein synthesis, which is one of the most important parameters of cell renewal, with excellent sensitivity.
使用磺酰罗丹明B(SRB)开展了一项比色化学敏感性试验,SRB可对细胞合成的蛋白质进行染色,作为终点标志物。对4种培养的细胞系、9种连续移植到裸鼠体内的人肿瘤异种移植物以及14份新鲜手术标本进行了该检测。该检测的最佳条件为:96孔板中每孔3 - 5×10⁴个细胞、SRB浓度为4%以及孵育时间超过10分钟。当通过SRB检测评估丝裂霉素C、阿霉素、顺铂和5 - 氟尿嘧啶时,浓度 - 效应曲线在低浓度和高浓度平台之间显示出陡峭的斜率,这表明该检测具有出色的灵敏度,能够将药物的效果评估为“全或无”。尽管这种高灵敏度使得该检测对培养细胞系具有良好的可重复性,但SRB检测对体内人肿瘤异种移植物化学敏感性的预测率仅为63.9%。因此,由于SRB检测能够以出色的灵敏度直接评估细胞蛋白质合成(细胞更新的最重要参数之一),所以它被认为对于评估培养细胞的化学敏感性为全或无是有用的。