NADPH细胞色素P450还原酶激活的伊达比星在敏感和多药耐药MCF7乳腺癌细胞中诱导的DNA损伤

DNA damage induced by NADPH cytochrome P450 reductase-activated idarubicin in sensitive and multidrug resistant MCF7 breast cancer cells.

作者信息

Żwierełło Wojciech, Maruszewska Agnieszka, Nowak Robert, Kostrzewa-Nowak Dorota, Tarasiuk Jolanta

机构信息

Department of Biochemistry, Faculty of Biology, University of Szczecin, 3c Felczaka St., 71-412 Szczecin, Poland; Molecular Biology and Biotechnology Center, Faculty of Biology, University of Szczecin, 71-415 Szczecin, 13 Wąska St., Poland.

出版信息

Pharmacol Rep. 2017 Feb;69(1):185-195. doi: 10.1016/j.pharep.2016.10.002. Epub 2016 Oct 4.

DOI:10.1016/j.pharep.2016.10.002

PMID:27940401

Abstract

BACKGROUND

Idarubicin (IDA) is one of clinically important anticancer drugs belonging to the anthracycline antibiotic family. The aim of this study was to examine DNA damage induced by NADPH cytochrome P450 reductase (CPR)-activated IDA in human sensitive MCF7 and multidrug resistant MCF7/DOX (overexpressing P-gp) breast adenocarcinoma cells.

METHODS

The evaluation of DNA fragmentation caused by single strand breaks (SSB) and double strand breaks (DSB) was performed using terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) test. Additionally, DSB formation was examined using H2AX histone phosphorylation assays.

RESULTS

It was found that IDA alone and CPR-activated used at IC caused a higher level of DNA strand breaks in sensitive MCF7 cells detected by TUNEL assessments (p=0.0011 for IDA alone and p=0.0109 for IDA reductively activated, Kruskal-Wallis test) and γ-H2AX-positive staining (p=0.0003 for IDA alone and p=0.0193 for IDA reductively activated, Kruskal-Wallis test) than in multidrug resistant MCF7/DOX cells. However, no changes were observed in the percentage of TUNEL-positive and DSB-positive cells for MCF7 as well as MCF7/DOX cells in the case of IDA alone and the drug pretreated in the presence of the activating system.

CONCLUSIONS

The obtained results suggest that CPR-activation of IDA does not significantly change the cellular DNA damage response of studied sensitive MCF7 and multidrug resistant MCF7/DOX breast cancer cells, even if the results concerning the interaction of IDA undergoing CPR activation with naked DNA showed the important differences in comparison with the drug alone (non-activated).

摘要

背景

伊达比星（IDA）是临床上重要的抗癌药物之一，属于蒽环类抗生素家族。本研究的目的是检测烟酰胺腺嘌呤二核苷酸磷酸细胞色素P450还原酶（CPR）激活的IDA在人敏感MCF7和多药耐药MCF7/DOX（过表达P-糖蛋白）乳腺腺癌细胞中诱导的DNA损伤。

方法

使用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）试验评估单链断裂（SSB）和双链断裂（DSB）引起的DNA片段化。此外，使用H2AX组蛋白磷酸化分析检测DSB的形成。

结果

发现单独使用IDA和在IC下使用CPR激活的IDA在敏感MCF7细胞中导致的DNA链断裂水平高于多药耐药MCF7/DOX细胞，这通过TUNEL评估检测到（单独使用IDA时p=0.0011，还原激活的IDA时p=0.0109，Kruskal-Wallis检验）以及γ-H2AX阳性染色（单独使用IDA时p=0.0003，还原激活的IDA时p=0.0193，Kruskal-Wallis检验）。然而，对于单独使用IDA以及在激活系统存在下预处理的药物，MCF7和MCF7/DOX细胞中TUNEL阳性和DSB阳性细胞的百分比没有观察到变化。