Wilson J K, Sargent J M, Elgie A W, Hill J G, Taylor C G
Department of Gynaecology, Pembury Hospital, Kent, UK.
Br J Cancer. 1990 Aug;62(2):189-94. doi: 10.1038/bjc.1990.258.
We assess the feasibility of using the MTT assay as a measure of cell viability in chemosensitivity testing in ovarian malignancy. The assay utilises the conversion of the tetrazolium salt MTT to formazan by dehydrogenase enzymes in living cells. We show that the optical density of the formazan produced from MTT is directly proportional to the number of live cells tested. Optimum MTT conversion occurred after 4 h incubation and dimethyl sulphoxide was found to be the most suitable solvent for the formazan. Seventy-five samples of ascitic fluid and/or solid tumour were collected from 56 patients with FIGO stage III-IV ovarian adenocarcinoma. Malignant cell suspensions with a viability greater than 75% were prepared from 95% of ascitic fluid and 75% of biopsy samples by simple techniques. The effect of cytotoxic drugs was assessed in 91% of patients included in the study. Variation in drug effect between patients was evident following a 48 h incubation period and was reproducible. Overall platinum and anthraquinone analogues produced the greater effect but resistance did occur. Our results mirrored reported clinical response rates. Only one sample tested against chlorambucil showed any drug effect. As this assay produces results in a high percentage of tests and is rapid and simple it appears suitable for prospective clinical trials to correlate the in vitro results with in vivo response.
我们评估了在卵巢恶性肿瘤的化疗敏感性测试中使用MTT法作为细胞活力检测方法的可行性。该检测方法利用活细胞中的脱氢酶将四唑盐MTT转化为甲臜。我们发现，MTT产生的甲臜的光密度与所检测的活细胞数量直接成正比。孵育4小时后MTT转化达到最佳，并且发现二甲基亚砜是最适合甲臜的溶剂。从56例国际妇产科联盟（FIGO）III-IV期卵巢腺癌患者中收集了75份腹水和/或实体瘤样本。通过简单技术，从95%的腹水样本和75%的活检样本中制备出活力大于75%的恶性细胞悬液。在纳入研究的91%的患者中评估了细胞毒性药物的效果。经过48小时的孵育期后，患者之间药物效果的差异明显且可重复。总体而言，铂类和蒽醌类类似物产生的效果更佳，但确实出现了耐药情况。我们的结果与报道的临床缓解率相符。仅一份针对苯丁酸氮芥检测的样本显示出任何药物效果。由于该检测方法在高比例的测试中能产生结果，且快速简便，因此似乎适用于前瞻性临床试验，以将体外结果与体内反应相关联。
