Csoka K, Larsson R, Tholander B, Gerdin E, de la Torre M, Nygren P
Division of Clinical Pharmacology, University Hospital, Uppsala University, Sweden.
Gynecol Oncol. 1994 Aug;54(2):163-70. doi: 10.1006/gyno.1994.1187.
The automated fluorometric microculture cytotoxicity assay (FMCA) is based on the measurement of fluorescence generated from cellular hydrolysis of fluorescein diacetate (FDA) to fluorescein by viable cells after a 72-hr culture period in microtiter plates. The FMCA was adopted for chemosensitivity testing of tumor cells from patients with ovarian carcinoma. Thirty-seven samples of solid tumors and malignant effusions were obtained from 35 patients at diagnosis or relapse. Tumor cells from solid samples and effusions were prepared by enzymatic digestion and centrifugation, respectively, followed by Percoll or Ficoll purification. The fluorescence was proportional to the number of cells/well and considerably higher in tumor cells than in contaminating normal cells. The effect of up to 19 cytotoxic drugs was successfully assessed in 70% of the samples and there was a good correlation between drug sensitivity data reported by the FMCA and the DiSC assay performed in parallel. The overall drug sensitivity pattern in vitro corresponded well to the clinical experience. The effect of cisplatin varied considerably between patients and resistance was found also in cases not previously exposed to cytotoxic drugs. The FMCA is a rapid and simple method that seems to report clinically relevant cytotoxic drug sensitivity data in ovarian carcinomas. In the future, this method may contribute to optimizing chemotherapy by assisting in individualized drug selection and new drug development.
自动荧光微量培养细胞毒性测定法（FMCA）基于在微量滴定板中培养72小时后，活细胞将二醋酸荧光素（FDA）水解为荧光素所产生的荧光测量。FMCA被用于卵巢癌患者肿瘤细胞的化疗敏感性测试。在诊断或复发时，从35例患者中获取了37份实体瘤和恶性积液样本。实体样本和积液中的肿瘤细胞分别通过酶消化和离心制备，随后进行Percoll或Ficoll纯化。荧光与每孔细胞数量成正比，且在肿瘤细胞中比在污染的正常细胞中高得多。在70%的样本中成功评估了多达19种细胞毒性药物的效果，FMCA报告的药物敏感性数据与同时进行的DiSC测定法之间具有良好的相关性。体外总体药物敏感性模式与临床经验相符。顺铂的效果在患者之间差异很大，并且在以前未接触过细胞毒性药物的病例中也发现了耐药性。FMCA是一种快速简单的方法，似乎能报告卵巢癌中与临床相关的细胞毒性药物敏感性数据。未来，这种方法可能通过协助个体化药物选择和新药开发，为优化化疗做出贡献。
