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双去甲氧基姜黄素诱导NCI-H460人肺癌细胞中的DNA损伤并抑制与DNA修复相关的蛋白表达。

Bisdemethoxycurcumin induces DNA damage and inhibits DNA repair associated protein expressions in NCI-H460 human lung cancer cells.

作者信息

Yu Chien-Chih, Yang Su-Tso, Huang Wen-Wen, Peng Shu-Fen, Huang An-Cheng, Tang Nou-Ying, Liu Hsin-Chung, Yang Mei-Due, Lai Kuang-Chi, Chung Jing-Gung

机构信息

School of Pharmacy, China Medical University, Taichung, 404, Taiwan.

Department of Radiology, China Medical University Hospital, Taichung, 404, Taiwan.

出版信息

Environ Toxicol. 2016 Dec;31(12):1859-1868. doi: 10.1002/tox.22187. Epub 2015 Aug 30.

Abstract

Nonsmall cell lung carcinoma (NSCLC) is a devastating primary lung tumor resistant to conventional therapies. Bisdemethoxycurcumin (BDMC) is one of curcumin derivate from Turmeric and has been shown to induce NSCLC cell death. Although there is one report to show BDMC induced DNA double strand breaks, however, no available information to show BDMC induced DNA damage action with inhibited DNA repair protein in lung cancer cells in detail. In this study, we tested BDMC-induced DNA damage and condensation in NCI-H460 cells by using Comet assay and DAPI staining examinations, respectively and we found BDMC induced DNA damage and condension. Western blotting was used to examine the effects of BDMC on protein expression associated with DNA damage and repair and results indicated that BDMC suppressed the protein levels associated with DNA damage and repair, such as 14-3-3σ (an important checkpoint keeper of DDR), O6-methylguanine-DNA methyltransferase, DNA repair proteins breast cancer 1, early onset, mediator of DNA damage checkpoint 1 but activate phosphorylated p53 and p-H2A.X (phospho Ser140) in NCI-H460 cells. Confocal laser systems microscopy was used for examining the protein translocation and results show that BDMC increased the translocation of p-p53 and p-H2A.X (phospho Ser140) from cytosol to nuclei in NCI-H460 cells. In conclusion, BDMC induced DNA damage and condension and affect DNA repair proteins in NCI-H460 cells in vitro. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1859-1868, 2016.

摘要

非小细胞肺癌(NSCLC)是一种对传统疗法具有抗性的毁灭性原发性肺癌。双去甲氧基姜黄素(BDMC)是姜黄中的一种姜黄素衍生物,已被证明可诱导NSCLC细胞死亡。尽管有一份报告显示BDMC可诱导DNA双链断裂,然而,尚无详细信息表明BDMC在肺癌细胞中通过抑制DNA修复蛋白诱导DNA损伤作用。在本研究中,我们分别使用彗星试验和DAPI染色检查测试了BDMC在NCI-H460细胞中诱导的DNA损伤和凝聚,我们发现BDMC可诱导DNA损伤和凝聚。蛋白质印迹法用于检测BDMC对与DNA损伤和修复相关的蛋白质表达的影响,结果表明BDMC可抑制与DNA损伤和修复相关的蛋白质水平,如14-3-3σ(DNA损伤反应的重要检查点维持蛋白)、O6-甲基鸟嘌呤-DNA甲基转移酶、DNA修复蛋白乳腺癌1、早发性、DNA损伤检查点1的介质,但可激活NCI-H460细胞中磷酸化的p53和p-H2A.X(磷酸化丝氨酸140)。共聚焦激光系统显微镜用于检测蛋白质转位,结果显示BDMC可增加NCI-H460细胞中p-p53和p-H2A.X(磷酸化丝氨酸140)从细胞质到细胞核的转位。总之,BDMC在体外可诱导NCI-H460细胞中的DNA损伤和凝聚,并影响DNA修复蛋白。©2015威利期刊公司。《环境毒理学》31: 1859-1868, 2016。

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