Zhang Guo-Jun, Chen Tsing-Bau, Bednar Bohumil, Connolly Brett M, Hargreaves Richard, Sur Cyrille, Williams David L
Imaging Department, MRL, Merck and Co., Inc., West Point, PA 19486, USA.
Neoplasia. 2007 Aug;9(8):652-61. doi: 10.1593/neo.07421.
The in vivo hollow fiber assay, in which semipermeable hollow fibers filled with tumor cells, are implanted into animals, was originally developed to screen for anticancer compounds before assessment in more complex tumor models. To enhance screening and evaluation of anticancer drugs, we have applied optical imaging technology to this assay. To demonstrate that tumor cells inside hollow fibers can communicate with the host mice, we have used fluorescence imaging in vivo and CD31 immunostaining ex vivo to show that angiogenesis occurs around cell-filled hollow fibers by 2 weeks after subcutaneous implantation. Bioluminescence imaging has been used to follow the number of luciferase-expressing tumor cells within implanted hollow fibers; proliferation of those cells was found to be significantly inhibited by docetaxel or irinotecan. We also used bioluminescence imaging of hollow fibers to monitor the nuclear factor kappaB (NFkappaB) pathway in vivo; NFkappaB activation by lipopolysaccharide and tumor necrosis factor-alpha was evaluated in tumor cell lines genetically engineered to express luciferase controlled by an NFkappaB-responsive element. These results demonstrate that optical imaging of hollow fibers containing reporter tumor cells can be used for the rapid and accurate evaluation of antitumor activities of anticancer drugs and for measurement of molecular pathways.
体内中空纤维试验是将填充有肿瘤细胞的半透性中空纤维植入动物体内，该试验最初是为了在更复杂的肿瘤模型评估之前筛选抗癌化合物而开发的。为了加强抗癌药物的筛选和评估，我们已将光学成像技术应用于该试验。为了证明中空纤维内的肿瘤细胞可与宿主小鼠进行交流，我们在体内使用了荧光成像，在体外使用了CD31免疫染色，以显示皮下植入后2周，充满细胞的中空纤维周围发生了血管生成。生物发光成像已用于追踪植入的中空纤维内表达荧光素酶的肿瘤细胞数量；发现多西他赛或伊立替康可显著抑制这些细胞的增殖。我们还使用中空纤维的生物发光成像在体内监测核因子κB（NFκB）信号通路；在经过基因工程改造以表达受NFκB反应元件控制的荧光素酶的肿瘤细胞系中，评估了脂多糖和肿瘤坏死因子-α对NFκB的激活作用。这些结果表明，含有报告基因肿瘤细胞的中空纤维的光学成像可用于快速、准确地评估抗癌药物的抗肿瘤活性以及测量分子信号通路。
