Náray-Fejes-Tóth A, Fejes-Tóth G, Volk K A, Stokes J B
Department of Physiology, Dartmouth Medical School, Lebanon, NH 03756-0001, USA.
J Steroid Biochem Mol Biol. 2000 Dec 1;75(1):51-6. doi: 10.1016/s0960-0760(00)00136-9.
Serum- and glucocorticoid-induced kinase (sgk) is transcriptionally regulated by corticosteroids in several cell types. Recent findings suggest that sgk is an important gene in the early action of corticosteroids on epithelial sodium reabsorption. Surprisingly, the human sgk was reported not to be transcriptionally regulated by corticosteroids in a hepatoma cell line, and thus far no glucocorticoid response element has been identified in the human SGK gene. Since humans clearly respond to both aldosterone and glucocorticoids in cells where sgk action seems to be important, in this study we determined sgk mRNA levels following dexamethasone treatment for various duration in five human cell lines. These cell lines included epithelial cells (H441, T84 and HT29) and lymphoid/monocyte (U937 and THP-1) lines. Using quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), we found that sgk mRNA levels are markedly induced by glucocorticoids in all of the five cell lines studied. Time course analyses revealed that sgk mRNA levels are elevated as early as 30 min after addition of the glucocorticoid, and remain elevated for several hours. Northern analysis in H441 cells confirmed that sgk is an early induced gene. The induction of sgk by dexamethasone was unaffected by cycloheximide, indicating that it does not require de novo protein synthesis. These results indicate that the human sgk, just like its counterparts in other species, is a primary glucocorticoid-induced gene.
血清和糖皮质激素诱导激酶(sgk)在多种细胞类型中受糖皮质激素的转录调控。最近的研究结果表明,sgk是糖皮质激素对上皮钠重吸收早期作用中的一个重要基因。令人惊讶的是,据报道人类sgk在肝癌细胞系中不受糖皮质激素的转录调控,而且迄今为止在人类SGK基因中尚未鉴定出糖皮质激素反应元件。由于人类在sgk作用似乎很重要的细胞中对醛固酮和糖皮质激素都有明显反应,因此在本研究中,我们测定了五种人类细胞系经地塞米松处理不同时间后的sgk mRNA水平。这些细胞系包括上皮细胞系(H441、T84和HT29)以及淋巴/单核细胞系(U937和THP-1)。使用定量逆转录聚合酶链反应(RT-PCR),我们发现在所研究的所有五种细胞系中,糖皮质激素均能显著诱导sgk mRNA水平升高。时间进程分析显示,早在添加糖皮质激素后30分钟,sgk mRNA水平就开始升高,并持续升高数小时。对H441细胞进行的Northern分析证实sgk是一个早期诱导基因。地塞米松对sgk的诱导不受放线菌酮的影响,这表明其诱导不需要从头合成蛋白质。这些结果表明,人类sgk与其他物种的同类基因一样,是一个主要的糖皮质激素诱导基因。