培养的人外周血白细胞对抗肿瘤药物5,6-二甲基呫吨酮-4-乙酸（NSC 640488）产生肿瘤坏死因子-α。

Production of tumour necrosis factor-alpha by cultured human peripheral blood leucocytes in response to the anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid (NSC 640488).

作者信息

Philpott M, Joseph W R, Crosier K E, Baguley B C, Ching L M

机构信息

Cancer Research Laboratory, University of Auckland School of Medicine, New Zealand.

出版信息

Br J Cancer. 1997;76(12):1586-91. doi: 10.1038/bjc.1997.601.

DOI:10.1038/bjc.1997.601

PMID:9413946

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2228206/

Abstract

The investigative anti-tumour agent 5,6-dimethylxanthenonone-4-acetic acid (DMXAA, NSC 640488), developed in this laboratory as an improved analogue of flavone acetic acid (FAA, NSC 347512), is currently in clinical trial. The ability of DMXAA to up-regulate tumour necrosis factor (TNF) mRNA and protein synthesis in cultured human peripheral blood leucocytes (HPBLs) has been investigated and compared with that of flavone acetic acid (FAA) and of bacterial lipopolysaccharide (LPS). Human peripheral blood leucocytes were isolated from buffy coats obtained from a blood transfusion centre and also from blood samples from laboratory volunteers. At a concentration of 400 microg ml(-1) and an incubation time of 2 h, DMXAA up-regulated mRNA synthesis in six of eight individuals tested, as measured by Northern blotting. The degree of up-regulation varied in different individuals from one to nine times that of control levels. In contrast, FAA caused no induction above that of control levels and in some cases suppressed expression relative to controls, extending previous data that DMXAA but not FAA up-regulates TNF mRNA in the human HL-60 tumour cell line. At the same concentration but with longer incubation times (6-12 h), DMXAA induced increases in TNF protein in 11 of 15 samples of HPBLs from buffy coats and also in 11 of 15 samples of HPBLs from volunteers, as measured by cytotoxicity assays with L929 cells. FAA caused no increase in TNF protein, while LPS induced TNF to approximately 20-fold higher levels than did DMXAA. Considerable heterogeneity of response was observed with both sources of HPBLs, and there was little or no correlation between the extent of TNF induction by DMXAA and LPS in individual samples. In vitro analysis of the response of human peripheral blood leucocytes to DMXAA may be a useful test in clinical trials of agents such as DMXAA.

摘要

本实验室研发的研究性抗肿瘤药物5,6-二甲基呫吨酮-4-乙酸（DMXAA，NSC 640488），是黄酮乙酸（FAA，NSC 347512）的改良类似物，目前正处于临床试验阶段。已对DMXAA上调培养的人外周血白细胞（HPBL）中肿瘤坏死因子（TNF）mRNA和蛋白质合成的能力进行了研究，并与黄酮乙酸（FAA）和细菌脂多糖（LPS）的能力进行了比较。人外周血白细胞从输血中心获得的血沉棕黄层以及实验室志愿者的血样中分离得到。在浓度为400 μg ml⁻¹且孵育时间为2小时的条件下，通过Northern印迹法检测，DMXAA在8名受试个体中的6名中上调了mRNA合成。上调程度在不同个体中有所不同，为对照水平的1至9倍。相比之下，FAA未引起高于对照水平的诱导，在某些情况下相对于对照抑制了表达，这扩展了之前关于DMXAA而非FAA在人HL-60肿瘤细胞系中上调TNF mRNA的数据。在相同浓度但孵育时间更长（6 - 12小时）的情况下，通过用L929细胞进行细胞毒性测定，DMXAA在来自血沉棕黄层的15份HPBL样本中的11份以及来自志愿者的15份HPBL样本中的11份中诱导了TNF蛋白增加。FAA未引起TNF蛋白增加，而LPS诱导的TNF水平比DMXAA高约20倍。在两种HPBL来源中均观察到反应的显著异质性，并且在单个样本中DMXAA和LPS诱导TNF的程度之间几乎没有相关性。对人外周血白细胞对DMXAA反应的体外分析可能是DMXAA等药物临床试验中的一项有用测试。