角质形成细胞中的腺苷酸环化酶：FRSK细胞表达I型、II型、III型、IV型、VI型和VIII型，并且在同工酶表达未改变的情况下，1,25(OH)₂D₃、视黄酸和佛波酯增强了福斯高林诱导的环磷酸腺苷积累。

Adenylate cyclases in keratinocytes: FRSK cells express types I, II, III, IV, VI and VIII, and 1,25(OH)2D3, retinoic acid and TPA augment forskolin-induced cyclic AMP accumulation in the absence of altered isozyme expression.

作者信息

Takahashi H, Kinouchi M, Tamura T, Ishida-Yamamoto A, Iizuka H

机构信息

Department of Dermatology, Asahikawa Medical College, Japan.

出版信息

Arch Dermatol Res. 1998 Aug;290(8):407-12. doi: 10.1007/s004030050327.

DOI:10.1007/s004030050327

PMID:9763301

Abstract

Molecular cloning analysis has detected at least nine adenylate cyclase isozymes in mammalian tissues. Using fetal rat skin keratinocytes (FRSK), we investigated adenylate cyclase expression and its modulation by 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), a retinoid (Ro10-1670), and 12-O-tetradecanoylphorbol-13-acetate (TPA). Reverse transcription polymerase chain reaction (RT-PCR) indicated that FRSK contain adenylate cyclases I, II, III, IV, VI and VIII. Treatment with 1,25(OH)2D3 (1 x 10(-7) M), Ro10-1670 (1 x 10(-6) M), and TPA (100 ng/ml) resulted in increased forskolin-induced cyclic AMP accumulation by FRSK cells and normal human keratinocytes (NHK). Quantitative RT-PCR and Western blot analysis, however, detected no alteration in mRNA and protein levels of each adenylate cyclase isozyme for at least 48 h. These results indicate that FRSK contain at least six (I, II, III, IV, VI and VIII) adenylate cyclase isozyme mRNAs, suggesting a complex regulatory mechanism of cyclic AMP generation in keratinocytes. Although 1,25(OH)2D3, Ro10-1670, and TPA augmented forskolin-induced cyclic AMP accumulation, they do not seem to affect the expression of specific adenylate cyclase isozymes by FRSK cells.

摘要

分子克隆分析已在哺乳动物组织中检测到至少九种腺苷酸环化酶同工酶。我们使用胎鼠皮肤角质形成细胞（FRSK），研究了腺苷酸环化酶的表达及其受1,25-二羟基维生素D3（1,25(OH)2D3）、一种类视黄醇（Ro10-1670）和12-O-十四烷酰佛波醇-13-乙酸酯（TPA）的调节情况。逆转录聚合酶链反应（RT-PCR）表明，FRSK含有腺苷酸环化酶I、II、III、IV、VI和VIII。用1,25(OH)2D3（1×10−7 M）、Ro10-1670（1×10−6 M）和TPA（100 ng/ml）处理导致FRSK细胞和正常人角质形成细胞（NHK）中福斯高林诱导的环磷酸腺苷（cAMP）积累增加。然而，定量RT-PCR和蛋白质印迹分析在至少48小时内未检测到每种腺苷酸环化酶同工酶的mRNA和蛋白质水平有变化。这些结果表明，FRSK含有至少六种（I、II、III、IV、VI和VIII）腺苷酸环化酶同工酶mRNA，提示角质形成细胞中环磷酸腺苷生成存在复杂的调节机制。尽管1,25(OH)2D3、Ro10-1670和TPA增强了福斯高林诱导的环磷酸腺苷积累，但它们似乎不影响FRSK细胞中特定腺苷酸环化酶同工酶的表达。