DNA damage response induced by Etoposide promotes steroidogenesis via GADD45A in cultured adrenal cells.
DNA damage response induced by Etoposide promotes steroidogenesis via GADD45A in cultured adrenal cells.
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DOI：
10.1038/s41598-018-27938-5
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发表时间：
2018-06-25
影响因子：
4.6
通讯作者：
Okazaki T
中科院分区：
文献类型：
--
作者：
Tamamori-Adachi M;Koga A;Susa T;Fujii H;Tsuchiya M;Okinaga H;Hisaki H;Iizuka M;Kitajima S;Okazaki T
肾上腺皮质中促肾上腺皮质激素(ACTH)通过环磷酸腺苷(CAMP)/蛋白激酶A(PKA)途径调节糖皮质激素的产生，但与衰老相关的类固醇合成的变化尚不清楚。在这项研究中，我们证明了非ACTH介导的遗传毒性应激在人肾上腺皮质H295R细胞中诱导了细胞自主的类固醇合成。小剂量依托泊苷(EP)作为一种遗传毒性应激诱导DNA损伤，导致细胞衰老。我们发现，用γ损伤细胞的标志物DNAH_2AX染色的细胞中，类固醇合成被促进。在应激相关和P53诱导基因中，GADD45A和类固醇生成相关基因的表达显著上调。免疫荧光分析显示GADD45A在细胞核内聚集。培养上清液中的代谢产物分析表明，EP处理的细胞被诱导产生和分泌相当数量的糖皮质激素。用小干扰RNA敲除GADD45A可显著抑制EP诱导的类固醇生成相关基因表达上调和糖皮质激素生成。P38MAPK抑制剂，而不是PKA抑制剂，可抑制EP刺激的类固醇合成。这些结果表明，DNA损伤本身通过一个或多个史无前例的非ACTH途径促进类固醇合成。具体地说，GADD45A在EP刺激的遗传毒性应激引发的类固醇合成过程中起着至关重要的作用。我们的研究为肾上腺皮质类固醇生成的另一种机制提供了新的线索。
Glucocorticoid production is regulated by adrenocorticotropic hormone (ACTH) via the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) pathway in the adrenal cortex, but the changes in steroidogenesis associated with aging are unknown. In this study, we show that cell-autonomous steroidogenesis is induced by non-ACTH- mediated genotoxic stress in human adrenocortical H295R cells. Low-dose etoposide (EP) was used to induce DNA damage as a genotoxic stress, leading to cellular senescence. We found that steroidogenesis was promoted in cells stained with γH2AX, a marker of DNA damaged cells. Among stress-associated and p53-inducible genes, the expression of GADD45A and steroidogenesis-related genes was significantly upregulated. Immunofluorescence analysis revealed that GADD45A accumulated in the nuclei. Metabolite assay using cultured media showed that EP-treated cells were induced to produce and secrete considerable amounts of glucocorticoid. Knockdown of GADD45A using small interfering RNA markedly inhibited the EP-induced upregulation of steroidogenesis-related gene expression, and glucocorticoid production. A p38MAPK inhibitor, but not a PKA inhibitor, suppressed EP-stimulated steroidogenesis. These results suggest that DNA damage itself promotes steroidogenesis via one or more unprecedented non-ACTH-mediated pathway. Specifically, GADD45A plays a crucial role in the steroidogenic processes triggered by EP-stimulated genotoxic stress. Our study sheds new light on an alternate mechanism of steroidogenesis in the adrenal cortex.
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影响因子：
3.9
作者：
Cerquetti, L.;Bucci, B.;Stigliano, A.
通讯作者：
Stigliano, A.
DOI：
10.1074/jbc.m116.740308
发表时间：
2016-08-19
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Bullard SA;Seo S;Schilling B;Dyle MC;Dierdorff JM;Ebert SM;DeLau AD;Gibson BW;Adams CM
通讯作者：
Adams CM
影响因子：
10.5
作者：
Brancho, D;Tanaka, N;Davis, RJ
通讯作者：
Davis, RJ
影响因子：
4.2
作者：
HEUSER, IJ;GOTTHARDT, U;HOLSBOER, F
通讯作者：
HOLSBOER, F
影响因子：
4.8
作者：
Connolly, Fiona;Rae, Michael T.;Duncan, W. Colin
通讯作者：
Duncan, W. Colin