Chemical ionization mass spectrometric determination of acrolein in human breast cancer cells

被引:38
作者
Kato, S
Post, GC
Bierbaum, VM
Koch, TH
机构
[1] Univ Colorado, Dept Chem & Biochem, Boulder, CO 80309 USA
[2] Univ Colorado, Ctr Canc, Denver, CO 80262 USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
chemical ionization mass spectrometry; acrolein; breast cancer cells; doxorubicin; drug resistance;
D O I
10.1006/abio.2002.5682
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A selected ion flow tube-chemical ionization mass spectrometric method is presented for the first determination of acrolein metabolically produced in biological tissues. Acrolein in aqueous samples (2.5 ml) is preconcentrated by distillation and directly analyzed using gas-phase proton transfer from H3O+. This method provides sensitive detection of acrolein with the method detection limit of 15 nM at the 99% confidence level. Detection is linear up to the highest concentration studied (13.5 muM, R-2 = 0.998). Acrolein levels are determined in doxorubicin-sensitive (MCF-7) and doxorubicin-resistant (MCF-7/Adr) human breast cancer cells in vitro. The intracellular acrolein concentrations differ insignificantly: 0.61 muM for sensitive cells and 0.54 muM for resistant cells. Treatment with a physiological concentration of doxorubicin (0.5 muM) for 24 h at 37degreesC increased acrolein levels by factors of 2.6 and 1.9 for MCF-7 and MCF-7/Adr cells, respectively. The differential enhancement observed is consistent with the lower levels of enzymes that neutralize oxidative stress in sensitive MCF-7 cells and overexpression of an active drug efflux pump P-170 glycoprotein in resistant MCF-7/Adr cells. (C) 2002 Elsevier Science (USA).
引用
收藏
页码:251 / 259
页数:9
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