经小动物内窥镜行肠黏膜下注射肿瘤细胞建立原位结直肠癌模型的技术探讨

目的:探讨使用活体小动物内窥镜进行原位注射人源性结肠癌细胞系建立原位结直肠癌模型的技术方法。方法:6~8周龄BALB/c雌性裸鼠20只，异氟醚气体持续麻醉，通过小动物结肠镜的引导，使用30G注射针将人结肠癌HCT-116细胞注射到裸鼠肠黏膜下，在肿瘤细胞注射3、7、10、15天进行小鼠内窥镜检查，观察裸鼠结直肠的成瘤情况。结果:利用小动物内窥镜行黏膜注射肿瘤细胞建立结直肠癌模型的技术成功率达100%。在注射肿瘤细胞时未出现穿孔，在小鼠内窥镜第7天随访时有1例裸鼠在肠镜通过肿瘤时出现肠壁穿孔。本研究中，没有出现因内窥镜注射导致的裸鼠死亡，所有19例存活裸鼠在第15天随访时均形成原位结直肠肿瘤，使用该方法建立裸鼠原位结直肠癌的建模成功率为95%。结论:通过小鼠内窥镜引导行肠黏膜下注射肿瘤细胞建立结直肠癌原位模型是一项快速而且有效的建模技术，该模型可更好地用于药物检测、基因功能评估和肿瘤转移。

Establishment of orthotopic colorectal cancer model by murine colonoscopy with submucosal injection of tumor cells in living mice

Objective:To explore the technique using murine colonoscopy to administer submucosal injection of tumor cells for the establishment of orthotopic colorectal cancer model.Methods:20 female BALB/c nude mice of 6~8 weeks were anesthetized continuously with isoflurane gas,given submocosal injection of human colon cancer HCT-116 cells through 30G entry needles under the guidance of murine colonoscopy,and examined by murine colonoscopy on the 3rd,7th,10th and 15th day after the injection to observe the tumor formation in the mouse colorectum.Results:The technique that using murine colonoscopy to administer submucosal injection of tumor cells for the establishment of orthotopic colorectal cancer model was performed with a success rate of 100%.During the injection of tumor cells,there was no perforation.However,1 nude mouse had intestinal wall perforation when the enteroscope passed the tumor on the 7th day after the injection during the follow-up.In this study,no nude mouse died due to endoscope injection.The tumor was formed in all 19 nude living mice on the 15th day after the injection,and the success rate for using this technique to establish orthotopic colorectal cancer model in nude mice was 95%.Conclusion:Using murine colonoscopy to administer submucosal injection of tumor cells for the establishment of orthotopic colorectal cancer model becomes a rapid and effective modeling technique.This model can be used to explore drug testing,gene function assessment and cancer metastasis.