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异硫氰酸苯己酯对K562/A02细胞系阿霉素耐药性的影响

[Effect of phenylhexyl isothiocyanate on adriamycin resistance of K562/A02 cell line].

作者信息

Yuang Peng, Chen Bao-An, Cheng Jian, Ma Xu-Dong, Liu De-Long, Lu Quan-Yi

机构信息

Department of Hematology, Zhongda Hospital, College of Clinical Medicine, Southeast University, Nanjing 210009, Jiangsu Province, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2009 Apr;17(2):352-7.

Abstract

This study was to investigate the effect of phenylhexyl isothiocyanate (PHI) on drug resistance and sensitivity on K562/A02 cell line to adriamycin (ADM) and to elucidate the possible mechanisms. The inhibition rates of ADM and PHI + ADM on growth of K562/A02 cell line were measured by MTT assay, and K562/A02 cell resistant multiple was calculated. The apoptosis rate of K562/A02 cell line, the changes of intracellular ADM concentrations and MRP1 protein level were detected by flow cytometry (FCM). Intracellular reoxidized GSH level was determined by spectrometric enzyme assay and MRP1 mRNA was assayed by semiquantitative RT-PCR before and after using PHI. The results indicated that the survival rate of K562/A02 cell line decreased with the increasing concentration of PHI. Apoptosis rate increased after treatment in combination with two above drugs, the changes of drug resistance multiple and intracellular ADM level had statistical significance between K562/A02 and K562 cells (p < 0.05), when the concentration of PHI was more than 20 micromol/L. Intracellular GSH level in K562/A02 cell line reduced 5% when 1 microg/ml ADM was used alone, and it increased slightly at first, then decreased when more than 10 micromol/L PHI was used. When more than 20 micromol/L PHI was used in combination with 1 microg/ml ADM, intracellular GSH level in K562/A02 cell line decreased progressively with increasing the concentration of PHI. The expressions of MRP1 mRNA and protein had no statistical significance between K562/A02 and K562 cells (p > 0.05) after or before PHI was used. It is concluded that the cyto-toxicity of PHI to K562/A02 cell line does not associate with the depletion of the intracellular GSH. PHI not only enhances the sensitivity of K562/A02 cell line to ADM, but also partially reverses effect of K562/A02 cell resistance to ADM. ADM combined with PHI can diminish side effect and dosage of ADM.

摘要

本研究旨在探讨异硫氰酸苯己酯(PHI)对K562/A02细胞株阿霉素(ADM)耐药性及敏感性的影响,并阐明其可能机制。采用MTT法检测ADM及PHI + ADM对K562/A02细胞株生长的抑制率,并计算K562/A02细胞耐药倍数。采用流式细胞术(FCM)检测K562/A02细胞株的凋亡率、细胞内ADM浓度变化及MRP1蛋白水平。使用PHI前后,通过分光光度酶法测定细胞内再氧化型谷胱甘肽(GSH)水平,采用半定量RT-PCR检测MRP1 mRNA。结果显示,K562/A02细胞株的存活率随PHI浓度升高而降低。两种药物联合处理后凋亡率升高,当PHI浓度大于20 μmol/L时,K562/A02与K562细胞之间的耐药倍数及细胞内ADM水平变化具有统计学意义(p < 0.05)。单独使用1 μg/ml ADM时,K562/A02细胞株细胞内GSH水平降低5%,使用10 μmol/L以上PHI时,细胞内GSH水平先略有升高,随后降低。当20 μmol/L以上PHI与1 μg/ml ADM联合使用时,K562/A02细胞株细胞内GSH水平随PHI浓度升高而逐渐降低。使用PHI前后,K562/A02与K562细胞之间MRP1 mRNA及蛋白表达无统计学意义(p > 0.05)。结论:PHI对K562/A02细胞株的细胞毒性与细胞内GSH消耗无关。PHI不仅增强K562/A02细胞株对ADM的敏感性,还部分逆转K562/A02细胞对ADM的耐药作用。ADM联合PHI可减少ADM的副作用及用量。

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