Characterization of retinoic acid-induced AP-1 activity in B16 mouse melanoma cells
Characterization of retinoic acid-induced AP-1 activity in B16 mouse melanoma cells
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DOI：
10.1074/jbc.272.19.12809
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发表时间：
1997-05-09
影响因子：
4.8
通讯作者：
Niles, RM
中科院分区：
文献类型：
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作者：
Desai, SH;Niles, RM
视黄酸（RA）诱导小鼠黑色素瘤细胞的分化，这伴随着蛋白激酶C α（PKC α）的增加以及核PKC α的选择性富集，用荧光素酶报告基因构建体瞬时转染B16细胞表明，RA诱导AP-1活性呈浓度依赖性增加，用二丁酸佛波酯（PDB）急性处理B16细胞（2 h）可使AP-1活性增加10倍。RA治疗没有改变Jun家族成员的表达;与此相反，急性PDB处理诱导了c-Fos的表达，而对c-Jun的影响不大。从B16细胞的核提取物和含有AP-1共有序列的寡核苷酸形成了五种DNA-蛋白复合物，RA处理的细胞中有几种复合物减少，相反，PKC的下调可抑制PDB和RA诱导的AP-1活性的急性升高，选择性PKC酶抑制剂双吲哚马来酰亚胺可降低PDB诱导的AP-1活性，但增强RA诱导的AP-1活性，这些结果与我们以前的研究表明，有趣的可能性，PKC蛋白，而不是酶的活性，可能需要RA诱导的AP-1活性。
Retinoic acid (RA) induces differentiation of Die mouse melanoma cells, which is accompanied by an increase in protein kinase C alpha (PKC alpha) as well as a selective enrichment of nuclear PKC alpha., We report here that RA also increases AP-1 activity in these cells, Transient transfection of B16 cells with luciferase reporter gene constructs indicated that RA induced a concentration-dependent increase in AP-1 activity, Acute treatment (2 h) of B16 cells with phorbol dibutyrate (PDB) increased AP-1 activity by 10-fold. RA treatment did not change the expression of Jun family members; however, it decreased the expression of c-Fos, In contrast acute PDB treatment induced c-Fos expression, while having little effect on c-Jun, Five DNA-protein complexes were formed with nuclear extracts from B16 cells and an oligonucleotide containing an AP-1 consensus sequence, Several complexes were decreased in cells treated with RA, Conversely, certain complexes were increased in cells acutely treated with PDB, The slowest migrating complexes were shown to contain Fos family members, Down-regulation of PKC inhibited both the acute PDB-induced and the RA-induced increase in AP-1 activity, The selective PKC enzyme inhibitor, bisindolylmaleimide, reduced PDB-stimulated AP-1 activity, but enhanced RA-induced AP-1 activity, These results together with our previous studies suggest the intriguing possibility that PKC protein, but not enzyme activity, may be required for RA-induced AP-1 activity.