摘要
目的在转录水平上分析某种信号引起细胞特定基因表达改变的量效关系。方法采用ＳＫＯ００７细胞，逐一对无损细胞核的制备、新生转录反应和［α３２Ｐ］标记新生ｍＲＮＡ的提取等关键步骤进行了方法学研究，并对我室研究白细胞介素（ＩＬ）６信号转导机制的常用细胞系ＩＬ６、ＩＬ６受体（Ｒ）α、ＩＬ６Ｒβ的表达进行了鉴定。结果１×１０５细胞即可得到阳性杂交信号，细胞数在５×１０５～１×１０７范围内，其杂交信号随细胞数的增加而增强，线性及重复性满意。常用细胞系除Ｍｏｌｔ４外，均有不同程度的ＩＬ６、ＩＬ６Ｒα和ＩＬ６Ｒβ表达。结论所建立的方法在受试细胞数仅为国外报道的十分之一时即可完成操作，且操作相对简单，易为人们接受。
Objective To establish nuclear runoff transcrption assay (NRTA) which may analyse specific gene expression of some cell lines in trancription levels aroused by the signal, so we can use NRTA in researching the role of IL 6 in multiple myeloma. Method By SKO 007 cells, technical parameters of NRTA during intact nuclei preparation, nuclear runoff transcription reaction, [α 32 P] mRNA purity, etc were studied. We established an improved newly NRTA and determined IL 6, IL 6R α and IL 6R β mRNA levels of some cell lines for researching signal transduction of IL 6. Results 1×10 5 cells were able to gain a positive hybridizational signal which is distinguishable. Hybridizational signals increased when cell quantity increased from 5×10 5 to 1×10 7. Its linear and repeativity were good. Five cell lines except Molt 4 exprese IL 6, IL 6R α, IL 6R β. Conclution The required cell quantity for test of the NRTA is only one tenth of reported openly. It is simple and easy.
