Molina A, Carpeaux R, Martial J A, Muller M
Laboratoire de Biologie Moléculaire et Génie Génétique, Université de Liège, Institut de Chimie B6, B-4000 Sart-Tilman, Belgium.
Toxicol In Vitro. 2002 Apr;16(2):201-7. doi: 10.1016/s0887-2333(01)00106-0.
We obtained a stable transformed fish (EPC) cell line containing a reporter gene under the control of the tilapia HSP70 promoter. Expression of the reporter gene, coding for a green fluorescent protein (GFP)-luciferase fusion protein, was assessed by measuring the luciferase enzymatic activity by luminometry and the GFP expression by fluorescence microscopy and flow cytometry. The clone was characterized for its capacity to respond to heat shock treatment. The results show high induction after 1 h at 37 degrees C of treatment, up to 500-fold. In addition, its convenience to detect a large range of cellular stressors was evaluated. We observed high induction when Cd2+, Zn2+, Hg2+ or Cu2+ was added, but not Pb2+. In addition, activation of the reporter gene was observed in the presence of other compounds such as acetyl chloride, tetrachlorophenol, chloroacetamide and sodium arsenite. In conclusion, this cell line can be used as a rapid, cheap and easy biological test to determine cellular stress induced by environmental pollutants, alone or in conjunction with other, more specific assays.
我们获得了一种稳定的转基因鱼类（EPC）细胞系，该细胞系含有一个受罗非鱼热休克蛋白70（HSP70）启动子控制的报告基因。通过发光法测量荧光素酶的酶活性以及通过荧光显微镜和流式细胞术检测绿色荧光蛋白（GFP）的表达，来评估编码GFP-荧光素酶融合蛋白的报告基因的表达。对该克隆进行了对热休克处理反应能力的表征。结果显示，在37摄氏度处理1小时后诱导率很高，高达500倍。此外，评估了其检测多种细胞应激源的便利性。当添加Cd2+、Zn2+、Hg2+或Cu2+时，我们观察到诱导率很高，但添加Pb2+时则不然。此外，在存在其他化合物如乙酰氯、四氯苯酚、氯乙酰胺和亚砷酸钠的情况下，观察到报告基因的激活。总之，该细胞系可作为一种快速、廉价且简便的生物学检测方法，用于单独或与其他更特异的检测方法联合，来确定环境污染物诱导的细胞应激。
