【摘要】 目的：了解不同刺激剂作用ＨＬ－６０细胞后对ＮＦ－ｋＢ活化的特点，为人类功能基因的筛选提供新的方法。方法：采用基因转染技术获得能稳定表达ＩｋＢａ－ＥＧＦＰ融合蛋白的ＨＬ－６０细胞系，定性和定量检测８种刺激剂作用于该细胞系后荧光强度的变化。结果：定性结果发现，在所用的８种刺激剂中，除ＩＬ－１ｒａ作用后的细胞荧光强度没有明显的变化外，其它７种刺激剂作用后都有不同程度的细胞荧光强度的降低，尤以ＰＨＡ作用后的变化更为典型。定量检测结果发现，稳定表达的ＨＬ－６０细胞自身的荧光强度，随时间延长即有降低，ＩＬ－１ｒａ作用后细胞荧光强度的降低同没有刺激时相比没有明显区别；其它７种刺激剂作用后均有明显的荧光强度降低，但它们作用后的特点并不相同。结论：该真核转染的细胞可用于ＮＫ－ｋＢ活化刺激剂的初步筛选，可以作为功能基因组研究中新基因功能初筛的一个平台技术。更多还原

【Abstract】 Objective:To understand the characterization of NE-KB activation stimulated with varying stimulators in RL-60,and provide a new approach for functional screening of novel genes. Methods: Qualitative observation and quantitative measurement of fluorescent intensity of HL-60 cells with pIkB-EGFP plasmid transfection stimulated by 100 ng/ml of 8 stimulators with fluorescent microscope and Polarstar fluorescent spectrometer. Results: It was found with fluorescent microscope that except IL- 1ra didn’ t make the fluoresent intensity decrease, the other 7 stimulators could decrease the fluorescent intensity of cells significantly, especially the fluorescent variety of the cells treated by PHA was more typical. The quantity results of fluorescent intensity ,were similar with the above qualitative results from florescent microscope. That was except the IL-1ira stimulation didn’t decrease the fluorescent intensity significantly, all the other 7 stimulators could decrease it, but the activated characterization of these stimulators was different. Conclusion:The above results indicated that, this IKB-EGFP stable expression cell could be employed to screen the NF-kB activation stimulators primarily, and it will become a bench-top technique in screen of novel gene functions.更多还原