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丁酸钠对人子宫内膜腺癌细胞剂量依赖性作用的流式细胞术及生化分析

Flow cytometric and biochemical analysis of dose-dependent effects of sodium butyrate on human endometrial adenocarcinoma cells.

作者信息

Saito S, Crissman H A, Nishijima M, Kagabu T, Nishiya I, Cram L S

机构信息

Department of Obsterics and Gynecology, Iwate Medical University, School of Medicine, Morioka, Japan.

出版信息

Cytometry. 1991;12(8):757-64. doi: 10.1002/cyto.990120810.

Abstract

Sodium butyrate (SB) treatment was previously shown to produce seven-fold increases in estrogen hormone receptor binding sites of human endometrial adenocarcinoma (IK) cells. Flow cytometric analysis and histone gel electrophoresis were used to examine cell cycle, cell metabolism, and nuclear histone fractions in IK cells treated with different concentrations of SB. SB-treated cells stained with fluorochromes specific for DNA, RNA, or general protein were analyzed by flow cytometry (FCM). Changes in accessibility to three DNA stains and gel electrophoresis were used to analyze rearrangements in chromatin structure. SB caused an accumulation of cells in the G1 phase and inhibited DNA synthesis, but not cellular levels of RNA and protein. Hoechst accessibility to A-T rich regions on DNA was dramatically increased after removal of SB. H1 histones were dephosphorylated and core histones were acetylated during SB-treatment. Information obtained in these studies may be useful for correlating cellular and biochemical events with SB-induced increases in nuclear steroid hormone binding sites.

摘要

先前的研究表明,丁酸钠(SB)处理可使人类子宫内膜腺癌(IK)细胞的雌激素激素受体结合位点增加7倍。采用流式细胞术分析和组蛋白凝胶电泳法,检测不同浓度SB处理的IK细胞的细胞周期、细胞代谢及核组蛋白组分。用特异性针对DNA、RNA或总蛋白的荧光染料对经SB处理的细胞进行染色,然后通过流式细胞术(FCM)进行分析。利用三种DNA染料的可及性变化和凝胶电泳分析染色质结构的重排。SB导致细胞在G1期积累并抑制DNA合成,但不影响细胞内RNA和蛋白质水平。去除SB后,DNA上富含A-T区域的Hoechst染料可及性显著增加。在SB处理过程中,H1组蛋白去磷酸化,核心组蛋白乙酰化。这些研究中获得的信息可能有助于将细胞和生化事件与SB诱导的核类固醇激素结合位点增加联系起来。

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