Jonsson B, Liminga G, Csoka K, Fridborg H, Dhar S, Nygren P, Larsson R
Division of Clinical Pharmacology, University Hospital, Uppsala University, Sweden.
Eur J Cancer. 1996 May;32A(5):883-7. doi: 10.1016/0959-8049(96)00015-9.
The aim of this study was to determine the in vitro cytotoxicity of calcein acetoxymethyl ester (Calcein/AM) on primary cultures derived from solid and haematological human tumours. Calcein/AM is a fluorescent dye that localises intracellularly after esterase-dependent cellular trapping and which has shown cytotoxic activity against various established human tumour cell lines at relatively low concentrations. The semi-automated fluorometric microculture cytotoxicity assay, based on the measurement of fluorescence generated from cellular hydrolysis of fluorescein diacetate to fluorescein, in microtitre plates was used for the evaluation of Calcein/AM activity in tumour cell suspensions from patients. The cytotoxicity was measured as a survival index (SI), defined as the fluorescence as a percentage of control cultures. A total of 163 evaluable samples from various tumours were tested with continuous drug exposure. The activity of Calcein/AM was compared with representatives of six major classes of standard chemotherapeutic drugs. Calcein/AM was found to induce concentration-dependent decreases in the SI of both haematological and solid tumour cells. The ratio of solid over haematological tumour activity increased at a rate that was concentration dependent. Although it was relatively less active than cisplatin against solid tumours, Calcein/AM showed higher solid tumour activity compared to leukaemic specific agents (cytarabine and amsacrine), vincristine and doxorubicin (Dox). Among the solid tumours tested, childhood tumours, non-small cell lung cancer and sarcomas were the most sensitive to Calcein/AM. The best correlation between SI values was seen between Calcein/AM and Dox, with weaker correlations to representatives of antimetabolites, platinum compounds, topoisomerase II inhibitors, tubulin interactive agents and alkylators. Non-cytotoxic concentrations of cyclosporin A significantly potentiated calcein-induced cytotoxicity. The results show that Calcein/AM is differentially active against haematological tumours, but with substantial activity against solid tumours. The drug may represent a new class of anticancer compound with a unique means of drug delivery.
本研究的目的是确定乙酰氧基甲基钙黄绿素（Calcein/AM）对源自人实体瘤和血液肿瘤的原代培养物的体外细胞毒性。Calcein/AM是一种荧光染料，在酯酶依赖性细胞捕获后定位于细胞内，并且在相对低的浓度下已显示出对各种已建立的人肿瘤细胞系的细胞毒性活性。基于在微量滴定板中测量由二乙酸荧光素细胞水解产生的荧光的半自动荧光微量培养细胞毒性测定法，用于评估患者肿瘤细胞悬液中Calcein/AM的活性。细胞毒性以存活指数（SI）来衡量，定义为荧光占对照培养物的百分比。对来自各种肿瘤的总共163个可评估样品进行了连续药物暴露测试。将Calcein/AM的活性与六种主要标准化疗药物的代表进行了比较。发现Calcein/AM可诱导血液肿瘤细胞和实体瘤细胞的SI呈浓度依赖性降低。实体瘤与血液肿瘤活性的比率以浓度依赖性速率增加。尽管与顺铂相比，Calcein/AM对实体瘤的活性相对较低，但与白血病特异性药物（阿糖胞苷和安吖啶）、长春新碱和多柔比星（Dox）相比，Calcein/AM显示出更高的实体瘤活性。在测试的实体瘤中，儿童肿瘤、非小细胞肺癌和肉瘤对Calcein/AM最敏感。Calcein/AM与Dox之间的SI值相关性最好，与抗代谢物、铂化合物、拓扑异构酶II抑制剂、微管蛋白相互作用剂和烷化剂的代表之间的相关性较弱。非细胞毒性浓度的环孢素A显著增强了钙黄绿素诱导的细胞毒性。结果表明，Calcein/AM对血液肿瘤具有不同的活性，但对实体瘤具有显著活性。该药物可能代表一类具有独特给药方式的新型抗癌化合物。
