Melding a New 3-Dimensional Agarose Colony Assay with the Emax Model to Determine the Effects of Drug Combinations on Cancer Cells
Melding a New 3-Dimensional Agarose Colony Assay with the Emax Model to Determine the Effects of Drug Combinations on Cancer Cells
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DOI：
10.1177/153303460900800210
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发表时间：
2009-04-01
影响因子：
2.8
通讯作者：
Levin, Victor A.
中科院分区：
文献类型：
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作者：
Kajiwara, Yoshinori;Panchabhai, Sonali;Levin, Victor A.
我们的研究目标有两个方面：（i）开发一种强大的3D集落测定方法，使用GelCount（TM）询问药物组合，以及（ii）开发可能在临床上用于治疗高级别胶质瘤的2种药物组合。我们使用三种胶质瘤细胞系（U251 MG、SNB 19和LNZ 308）和两种腺癌细胞系（MiaPaCa和SW 480）在双层琼脂糖培养物中作为集落生长。我们评估了二氟甲基鸟氨酸（DFMO），卡铂，伏立诺他（SAHA）和多西他赛的两种药物组合。为了分析抗肿瘤功效，我们使用GelCount（TM）测量每个平板中肿瘤集落体积（μ m（2）× OD）的曲线下面积。应用非线性剂量-反应E-max模型和基于Loewe加和性的相互作用指数计算两药协同、加和拮抗作用。对于胶质母细胞瘤细胞系，（I）卡铂后DFMO在2/3的细胞系中具有协同或加和作用，（ii）卡铂后SAHA在1个细胞系中具有协同作用，（iii）多西他赛之前的卡铂在2/3细胞系中是协同的，在第三细胞系中是部分加和的，（iv）多西他赛之前的SAHA在1/3细胞系中是协同的，（v）DFMO之前的多西他赛在3/3细胞系中是加和的或部分活性的，和（vi）DFMO加SAHA不活动，无论顺序如何。在MiaPaCA细胞系中，当DFMO跟随卡铂时，以及在短暴露时间内，当SAHA与卡铂组合时（无论顺序如何），发生协同作用。在SW 480细胞系中，仅在短时间暴露于卡铂和多西他赛后才发生协同作用; DFMO联合卡铂或多西他赛也可观察到相加和混合部分效应（无论顺序如何），卡铂先于DFMO，卡铂先于SAHA，多西他赛先于卡铂。总之，通过应用Gelcount（TM）自动计数和菌落大小测定以及使用E-max和Loewe模型来定义药物相互作用，我们可以可靠地将药物组合功效定义为对数剂量和药物暴露持续时间的函数。
The goal of our study was two-fold: (i) develop a robust 3D colony assay methodology to interrogate drug combinations using GelCount (TM) and (ii) to develop 2-drug combinations that might be useful in the clinic for the treatment of high-grade gliomas. We used three glioma cell lines (U251MG, SNB19, and LNZ308) and two adenocarcinoma cell lines (MiaPaCa and SW480) grown as colonies in a two-tiered agarose cultures. We evaluated two-drug combinations of difluoromethylomithine (DFMO), carboplatin, vorinostat (SAHA), and docetaxel. To analyze for antitumor efficacy we used GelCount (TM) to measure the area under the curve for tumor colony volumes (mu m(2) x OD) in each plate. The non-linear dose-response E-max model and the interaction index based on the Loewe additivity are applied to calculate two-drug synergy, additive, and antagonistic interactions.For glioblastoma cell lines, (I) carboplatin followed by DFMO was synergistic or additive in 2/3 cell lines, (ii) carboplatin before SAHA was synergistic in 1 cell line, (iii) carboplatin before docetaxel was synergistic in 2/3 cell lines and partially additive in the third, (iv) SAHA before docetaxel was synergistic in 1/3 cell lines, (v) docetaxel before DFMO was additive or partially active in 3/3 cell lines, and (vi) DFMO plus SAHA was inactive regardless of order. In the MiaPaCA cell line, synergy occurred when DFMO followed carboplatin and, at short exposure times, when SAHA was combined with carboplatin (regardless of order). In the SW480 cell line synergy occurred only in short exposures for carboplatin followed by docetaxel; additive and mixed partial effects were also seen with DFMO plus carboplatin or docetaxel (regardless of order), carboplatin before DFMO, carboplatin before SAHA, and docetaxel before carboplatin.In conclusion, by applying the Gelcount (TM) automated counting and sizing of colonies and the use of E-max and Loewe models to define drug interactions, we can reliably define drug combination efficacy as a function of log dose and duration of drug exposure.