利用荧光酶标仪通过Raji细胞中EB病毒早期抗原表达检测肿瘤启动子

Detection of tumor promoters by early antigen expression of EB virus in Raji cells using a fluorescence microplate-reader.

作者信息

Ohmori K, Miyazaki K, Umeda M

机构信息

Department of Food and Drug Science, Kanagawa Prefectural Public Health Laboratories, Yokohama, Japan.

出版信息

Cancer Lett. 1998 Oct 23;132(1-2):51-9. doi: 10.1016/s0304-3835(98)00159-1.

DOI:10.1016/s0304-3835(98)00159-1

PMID:10397453

Abstract

As an in vitro assay for possible tumor promoters, we designed a quantitative immunofluorometric method to detect Epstein-Barr virus early antigen (EBV-EA) expression in Raji cells. In this method, anti-EBV-EA monoclonal antibody, a fluorogenic substrate and a fluorescence microtiterplate-reader were employed. 12-O-Tetradecanoylphorbol-13-acetate (TPA) was shown to be a potent inducer. EBV-EA induction by TPA was related to the activation of protein kinase C and phospholipase A2. The chemicals that reacted positively were okadaic acid, diethylstilbestrol, progesterone, sodium phenobarbital, aldrin and dieldrin. Lithocholic acid, testosterone and DDT were equivocal in the present experiments. Eight other chemicals tested did not react.

摘要

作为一种检测潜在肿瘤促进剂的体外试验，我们设计了一种定量免疫荧光法来检测Raji细胞中EB病毒早期抗原（EBV-EA）的表达。在该方法中，使用了抗EBV-EA单克隆抗体、荧光底物和荧光微量滴定板读数器。结果表明，12-O-十四烷酰佛波醇-13-乙酸酯（TPA）是一种有效的诱导剂。TPA诱导EBV-EA与蛋白激酶C和磷脂酶A2的激活有关。呈阳性反应的化学物质有冈田酸、己烯雌酚、孕酮、苯巴比妥钠、艾氏剂和狄氏剂。在本实验中，石胆酸、睾酮和滴滴涕的反应不明确。其他八种测试化学物质没有反应。