Suggitt Marie, Swaine David J, Pettit George R, Bibby Michael C
Tom Connors Cancer Research Centre, University of Bradford, Bradford, United Kingdom.
Clin Cancer Res. 2004 Oct 1;10(19):6677-85. doi: 10.1158/1078-0432.CCR-04-0855.
The hollow fiber assay is used successfully as a routine in vivo screening model to quantitatively define anticancer activity by the National Cancer Institute. This study investigates whether the hollow fiber assay can be used as a short-term in vivo model to demonstrate specific pharmacodynamic end points, namely microtubule and cell cycle disruption.
The growth of A549 cells was characterized within hollow fibers over 5 days in vivo at both subcutaneous (s.c.) and intraperitoneal (i.p.) sites. Drugs were administered on day 4 (i.p.).
At 24 hours, cells were retrieved from fibers at both i.p. and s.c. sites of paclitaxel-treated (20 mg/kg) and combretastatin A1 phosphate-treated (150 mg/kg) mice. Cell cycle analysis after paclitaxel treatment revealed a mean G(2)-M phase population of 48.04% (i.p.) and 25.76% (s.c.) compared with vehicle group mice (6.78 and 5.56%, respectively; P = <0.001 and 0.005, respectively). Tumor cells retrieved from combretastatin A1 phosphate-treated mice had a mean G2-M phase population of 36.3% (i.p.) and 29.36% (s.c.) compared with cells retrieved from vehicle group mice (5.58 and 5.49%, respectively; P = <0.001). Using fluorescence and laser-confocal microscopy, paclitaxel was revealed to induce the formation of spindle asters and tubulin polymerization. Combretastatin A1 phosphate was shown to hold cells in mitosis. Changes in nuclear morphology were also observed.
These data demonstrate that the hollow fiber assay can be used as a short-term in vivo model for studying the pharmacodynamic effects of both standard and novel compounds on microtubules. Evidence has also been provided to support the routine use of the in vivo hollow fiber assay for demonstrating the mechanism of action of a drug.
中空纤维试验已成功用作美国国立癌症研究所定量定义抗癌活性的常规体内筛选模型。本研究调查中空纤维试验是否可作为短期体内模型来证明特定的药效学终点，即微管和细胞周期破坏。
在体内皮下（s.c.）和腹腔内（i.p.）部位，对A549细胞在中空纤维内5天的生长情况进行了表征。在第4天腹腔注射给药。
在24小时时，从紫杉醇处理（20mg/kg）和磷酸秋水仙素A1处理（150mg/kg）小鼠的腹腔内和皮下部位的纤维中回收细胞。紫杉醇处理后的细胞周期分析显示，与载体组小鼠相比，G(2)-M期群体平均为48.04%（腹腔内）和25.76%（皮下）（分别为6.78%和5.56%；P分别为<0.001和0.005）。与从载体组小鼠回收的细胞相比，从磷酸秋水仙素A1处理的小鼠回收的肿瘤细胞G2-M期群体平均为36.3%（腹腔内）和29.36%（皮下）（分别为5.58%和5.49%；P = <0.001）。使用荧光和激光共聚焦显微镜，发现紫杉醇诱导纺锤体星状体的形成和微管蛋白聚合。磷酸秋水仙素A1显示可使细胞停滞在有丝分裂期。还观察到核形态的变化。
这些数据表明，中空纤维试验可作为短期体内模型，用于研究标准化合物和新型化合物对微管的药效学作用。还提供了证据支持体内中空纤维试验用于证明药物作用机制的常规使用。
