MTT法抗肿瘤药敏试验影响因素的探讨

Study on the influencing factors of the MTT based tumor chemosensitivity assay in vitro

目的:对于采用MTT法进行抗肿瘤药物敏感试验过程中的多种影响因素进行分析,观察抗肿瘤药物包被板的稳定性。方法:分离人外周血单个核细胞(PBMC),用MTT法测定抗肿瘤药物敏感性,观察红细胞、血小板、3种DMSO及不同浓度MTT对实验结果的影响;监测肿瘤药物包被板的稳定性。结果:一定浓度范围的红细胞和血小板均可影响实验结果;进口和国产分析纯的DMSO与MTT的呈色反应是一致的,吸光度(A值)与MTT呈浓度依赖性;冷冻干燥的抗肿瘤药物包被板的稳定性优于未经冷冻干燥的药物包被板。结论:明确实验过程中多种影响因素,有助于规范MTT法体外抗肿瘤药敏试验。

Objective：To establish a standard process of measuring the anti-cancer drug sensitivity with MTT assay by analyzing the influencing factors of the chemosensitivity assay in vitro. Methods ： Some influencing factors of MTr assay in studying the sensitivity of human peripheral blood mononuclear ceils （PBMC） to anti-cancer drugs were observed, which contained red blood cells, platelets, three different kinds of DMSO and different concentration of MTF. Meanwhile we monitored the stability of tumor drug-coated plate. Results：The red blood cells and platelets may affect the results at a concentration certain range; analytical pure DMSO,both imported and domestic reagent showed the same color with MTT, the OD values of the reaction were dependent on MTT dose; The stability of freeze-drying the drug-coated plates were superior to non freeze-drying ones. Conclusion ： To clear and definite all kinds of influencing factors might helpful for standard MTT assay for drug sensitivity test in vitro.