胰腺癌神经浸润体外模型的构建与观察

Construction of models for perineural invasion in pancreatic cancer

目的构建胰腺癌神经浸润的体外模型，并对胰腺癌细胞的神经浸润现象进行初步观察。方法将胰腺癌Capan-2细胞与大鼠背根神经节（DRG）置于Matrigel基质胶中共培养，分别建立以观察细胞集落面积（模型1）和观察细胞迁移距离（模型2）为主的两种模型。倒置显微镜下观察神经突及细胞生长情况，利用图像分析软件Image pro plus对图片进行分析。结果共培养对神经突起生长无显著影响，无论是共培养还是单培养，DRG神经突均向四周呈放射状生长。培养模型中，Capan-2细胞朝向DRG方向生长迁移，接着包绕神经突呈纺锤状，并继续向DRG爬行。在模型1的共培养组，Capan-2细胞第5天时细胞集落面积为（309．28±19．11）斗m。，显著大于单培养模型的（208．57±7．94）μm。（P〈0．01）。在模型2的共培养组，Capan-2细胞第5天时向DRG方向迁移了（284．1±12．9）μm，而空白基质胶一侧，细胞迁移距离〈150μm，二者差异具有统计学意义（P〈0．01）。结论本实验成功构建了胰腺癌神经浸润体外模型，为后续研究打下了良好的实验基础。

Objective To establish in vitro model for perineural invasion （PNI） of pancreatic cancer, and observe the process of PNI. Methods Mouse dorsal root ganglia （DRG） and pancreatic cancer Capan-2 cell line were co-cultured in Matrigel matrix. Two models were constructed ： model 1 for observing areas of cell colonies and model 2 for observing distance of cell migration. Neurite outgrowth and cell colony growth were observed by invert microscope, images were analyzed by using Image pro plus software. Results Neurite outgrowth was not affected by co-culture. DRG neurite appeared radial growth under either co-culture or single culture. It was observed that Capan-2 cell could migrate to DRG, and grows around the nerve fiber as spindle- shaped, then continue to migrate to DRG. At the 5th day, in the co-culture group of model 1, the colony area was （ 309.28 ± 19.11 ） μm^2, which was significantly bigger than that in single culture model [ （ 208.57 ± 7.94） μm^2, P 〈0.01]. At the 5 th day, in the co-culture group of model 2, Capan-2 migrated （284.1 ±12.9） iJ.m to DRG, while in the side of blank Matrigel, cell migrated less than 150 μm, the difference was statistically significant （P 〈 0.01 ）. Conclusions The in vitro models for perineural invasion in pancreatic cancer were constructed successfully, which lay a good foundation for future studies.