Anderson M T, Wims L A, Morrison S L
Department of Microbiology, Columbia University, New York, NY.
Nucleic Acids Res. 1991 Dec;19(25):7235-41. doi: 10.1093/nar/19.25.7235.
In mature B cells the nuclear factor NF-kappa B which binds within the kappa enhancer is constitutively present in the nucleus. However, the lambda light chain producing myeloma J558L has been found to lack constitutively functional NF-kappa B. Deoxycholate released functional NF-kappa B from cytoplasmic extracts and functional NF-kappa B was present in J558L following cycloheximide but not phorbol ester treatment. J558L was also unable to respond to phorbol ester stimulation with synthesis of mRNA from the early response gene TIS11. J558L differs from S107, another myeloma which was found to be deficient in the synthesis of NF-kappa B but not in the activation of TIS11. Somatic cell hybrids were used to further define the defect in J558L; hybrids were made with the myelomas S107 and S194 and the pre-B cell line 70Z/3. In general, complementation of the defect in J558L was observed; however there was not a direct correlation between the levels of TIS11 mRNA and NF-kappa B expression in the somatic cell hybrids, suggesting that the pathways of activation of these genes, while possibly sharing common elements, are not identical. The defect in J558L was surprising given that it has frequently been used for the expression of transfected light chain genes.
在成熟B细胞中，结合于κ增强子内的核因子NF-κB组成性地存在于细胞核中。然而，已发现产生λ轻链的骨髓瘤J558L缺乏组成性功能型NF-κB。脱氧胆酸盐从细胞质提取物中释放出功能性NF-κB，在用环己酰亚胺处理而非佛波酯处理后，J558L中存在功能性NF-κB。J558L也无法通过早期反应基因TIS11的mRNA合成来响应佛波酯刺激。J558L与另一种骨髓瘤S107不同，后者被发现缺乏NF-κB的合成，但不缺乏TIS11的激活。体细胞杂交用于进一步确定J558L中的缺陷；与骨髓瘤S107和S194以及前B细胞系70Z/3进行杂交。总体而言，观察到J558L中的缺陷得到了互补；然而，体细胞杂交中TIS11 mRNA水平与NF-κB表达之间没有直接相关性，这表明这些基因的激活途径虽然可能共享共同元件，但并不相同。鉴于J558L经常用于转染轻链基因的表达，其缺陷令人惊讶。
