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μ类谷胱甘肽S-转移酶互补DNA的克隆及其在平滑肌肿瘤细胞系中糖皮质激素诱导性的表征。

Cloning of a mu-class glutathione S-transferase complementary DNA and characterization of its glucocorticoid inducibility in a smooth muscle tumor cell line.

作者信息

Norris J S, Schwartz D A, MacLeod S L, Fan W M, O'Brien T J, Harris S E, Trifiletti R, Cornett L E, Cooper T M, Levi W M

机构信息

Department of Medicine, Medical University of South Carolina, Charleston 29425-2229.

出版信息

Mol Endocrinol. 1991 Jul;5(7):979-86. doi: 10.1210/mend-5-7-979.

Abstract

A cDNA (designated hGSTYBX) encompassing the complete coding sequence of a hamster mu-class glutathione S-transferase (GST) subunit was cloned from a lambda ZAP library constructed with mRNA isolated from triamcinolone acetonide-treated smooth muscle tumor cells (DDT1 MF-2). Analysis of its nucleotide and deduced amino acid sequences demonstrated highest homology to the rat mu-class GST YB2 subunit. In proliferating subconfluent cells, in which constitutive expression of hGSTYBX mRNA was undetectable, glucocorticoid treatment induced hGSTYBX expression after a time lag of 3 h, and maximal induction occurred at 10 h. Nuclear run-on analysis showed that glucocorticoid induction resulted at least in part from an increased rate of transcription. Simultaneous treatment with glucocorticoid and cycloheximide prevented glucocorticoid induction, but had little effect on basal expression in confluent cells. In contrast, cycloheximide treatment 3 h after glucocorticoid treatment resulted in nearly full induction. These results taken together suggest that hGSTYBX induction may be a secondary glucocorticoid response.

摘要

从用曲安奈德处理的平滑肌肿瘤细胞(DDT1 MF-2)分离的mRNA构建的λZAP文库中克隆出一个包含仓鼠μ类谷胱甘肽S-转移酶(GST)亚基完整编码序列的cDNA(命名为hGSTYBX)。对其核苷酸和推导的氨基酸序列分析表明,它与大鼠μ类GST YB2亚基具有最高的同源性。在增殖的亚汇合细胞中,未检测到hGSTYBX mRNA的组成型表达,糖皮质激素处理在3小时的时间滞后诱导hGSTYBX表达,最大诱导发生在10小时。核转录分析表明,糖皮质激素诱导至少部分是由于转录速率增加。糖皮质激素和环己酰亚胺同时处理可阻止糖皮质激素诱导,但对汇合细胞的基础表达影响很小。相反,在糖皮质激素处理3小时后用环己酰亚胺处理导致几乎完全诱导。这些结果共同表明,hGSTYBX诱导可能是糖皮质激素的继发性反应。

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