Interaction of phorbol ester and adrenocorticotropin in the regulation of steroidogenic P450 genes in human fetal and adult adrenal cell cultures.
Interaction of phorbol ester and adrenocorticotropin in the regulation of steroidogenic P450 genes in human fetal and adult adrenal cell cultures.
复制标题
佛波酯和促肾上腺皮质激素在人胎儿和成人肾上腺细胞培养物中类固醇生成 P450 基因调节中的相互作用。
作者：
Vesa Ilvesmüki;R. Voutilainen
在人胎儿和成人肾上腺原代培养中研究了蛋白激酶c依赖机制在类固醇生成酶基因表达中的作用。细胞先培养7-10天，然后用ACTH或12- o -十四烷醇-13-醋酸酯（TPA）（一种蛋白激酶- c激活剂）刺激1-2天。提取胞质RNA，用32p标记的cDNA探针对P450scc（胆固醇侧链裂解酶/20,22-去糖化酶）、P450c17 （17 α -羟化酶/17,20-裂解酶）和P450c21（21-羟化酶）进行Northern和dot blotting分析；对于P450c11 （11 β -羟化酶/18-羟化酶/18-甲基氧化酶），使用30-mer寡核苷酸作为探针。ACTH （200 ng/ml）可使胎儿和成人培养物中所有甾体生成酶mrna的积累增加数倍。TPA以剂量依赖性的方式（0.01 ~ 100 ng/ml）抑制了这种积累，而非活性磷酯4 α -磷-12,13-二癸酸酯则没有作用。另一方面，在没有ACTH的情况下，TPA在成人培养中略微增加了所有类固醇性P450 mrna。在胎儿培养中，TPA略微增加了P450scc、P450c11和P450c21 mRNA的水平，而降低了P450c17 mRNA的水平。（Bu）2cAMP和霍乱毒素增加了促肾上腺皮质激素（ACTH）等甾体生成酶mrna。TPA下调（Bu）2cAMP-和霍乱毒素诱导的p450mrna的方式与acth诱导的mrna相同。在胎儿和成人培养中，促肾上腺皮质激素刺激的皮质醇、硫酸脱氢表雄酮和醛固酮的分泌均被TPA降低。在两种培养类型中，TPA都略微增加了基础类固醇产量。类固醇生成的变化与类固醇生成酶基因表达的变化密切相关。我们的研究结果表明，TPA对acth刺激的肾上腺类固醇生成的抑制作用是通过类固醇生成酶的mRNA水平介导的。因此，似乎蛋白激酶- c和camp依赖机制都参与了肾上腺皮质细胞中类固醇生成酶和激素产生的长期维持。
The role of protein kinase-C-dependent mechanisms in steroidogenic enzyme gene expression was studied in primary cultures of human fetal and adult adrenals. Cells were first cultured for 7-10 days and then stimulated with ACTH or 12-O-tetradecanoyl phorbol-13-acetate (TPA), a protein kinase-C activator, for 1-2 days. Cytoplasmic RNA was extracted and analyzed by Northern and dot blotting with 32P-labeled cDNA probes for P450scc (cholesterol side-chain cleavage enzyme/20,22-desmolase), P450c17 (17 alpha-hydroxylase/17,20-lyase), and P450c21 (21-hydroxylase); for P450c11 (11 beta-hydroxylase/18-hydroxylase/18-methyl oxidase), a 30-mer oligonucleotide was used as a probe. ACTH (200 ng/ml) increased the accumulation of all of the studied steroidogenic enzyme mRNAs in both fetal and adult cultures by several-fold. TPA inhibited this accumulation in a dose-dependent manner (0.01-100 ng/ml), whereas the inactive phorbol ester 4 alpha-phorbol-12,13-didecanoate was without effect. On the other hand, in the absence of ACTH, TPA slightly increased all steroidogenic P450 mRNAs in adult cultures. In fetal cultures TPA slightly increased P450scc, P450c11, and P450c21 mRNA levels, whereas it decreased P450c17 mRNA. (Bu)2cAMP and cholera toxin increased steroidogenic enzyme mRNAs such as ACTH. TPA down-regulated (Bu)2cAMP- and cholera toxin-induced P450mRNAs in the same way as ACTH-induced mRNAs. The secretion of ACTH-stimulated cortisol, dehydroepiandrosterone sulfate, and aldosterone was decreased by TPA in both fetal and adult cultures. The basal steroid production was slightly increased by TPA in both culture types. The changes in steroid production correlated well with the alterations in the steroidogenic enzyme gene expression. Our results show that the inhibitory effect of TPA on ACTH-stimulated adrenal steroidogenesis is mediated at the mRNA level of steroidogenic enzymes. Thus, it seems likely that both protein kinase-C- and cAMP-dependent mechanisms are involved in the long term maintenance of steroidogenic enzymes and hormone production in adrenocortical cells.
登录
查看更多内容
中小型卵泡猪颗粒细胞的类固醇生成：卵泡刺激激素、毛喉素和腺苷 3,5-环单磷酸与佛波酯的影响。
影响因子：
4.8
作者：
Hylka,VW;Kaki,MK;diZerega,GS
通讯作者：
diZerega,GS
佛波酯对培养的人胎儿肾上腺细胞中类固醇生成的作用。
影响因子：
2.1
作者：
Mason,JI;Carr,BR;Rainey,WE
通讯作者：
Rainey,WE
在培养的人肾上腺皮质细胞中，3,5-单磷酸腺苷和蛋白激酶 C 激活剂对 3β-羟基类固醇脱氢酶、17α-羟化酶和脱氢表雄酮磺基转移酶的双重调节。
影响因子：
4.8
作者：
McAllister,JM;Hornsby,PJ
通讯作者：
Hornsby,PJ
胎儿人肾上腺皮质的调节：促肾上腺皮质激素对胎儿和定形区细胞单层培养物的生长和功能的影响。
影响因子：
4.8
作者：
Simonian,MH;Gill,GN
通讯作者：
Gill,GN
硫酸甾醇在人类胎儿、无脑新生儿和成人肾上腺中的代谢。
DOI：
10.1210/jcem-55-4-671
发表时间：
1982
期刊：
The Journal of clinical endocrinology and metabolism
影响因子：
--
作者：
Korte,K;Hemsell,PG;Mason,JI
通讯作者：
Mason,JI