裸鼠胰腺癌高肝转移模型的建立

A pancreatic cancer model with high liver metastatic potential in nude mice

目的 建立高肝转移率的胰腺癌转移裸鼠模型，为抗胰腺癌肝转移药物的筛选和机理研究提供基础。方法 人胰腺癌MIA-PaCa2细胞株脾内接种形成肝转移病灶，经反复体外原代培养和体内接种后筛选出高转移亚型MIA-PaCa2-3，MIA-PaCa2-3细胞DAPI荧光标记后尾静脉注射建立裸鼠胰腺癌模型，小动物光学分子成像系统动态观察肝脏转移肿瘤的生长变化，荧光显微镜观察肝脏肿瘤细胞转移情况。采用Western-blot的方法比较MIA-PaCa2-3细胞及其源细胞MIA-PaCa2细胞转移部分相关基因的蛋白表达水平。结果 MIA-PaCa2-3细胞尾静脉注射裸鼠6～8周后，裸鼠活体肝脏区域及肝脏冰冻切片均可见明显弥漫性的荧光影像，肝脏转移率为80％（8／10），明显高于其源细胞MIA-PaCa2（2／10）；MIA-PaCa2-3细胞转移相关基因基质金属蛋白酶2（matrix metallo proteinase-2，MMP-2）、基质金属蛋白酶9（matrix metallo proteinase-9，MMP-9）、血管内皮细胞生长因子（Vascular endothelial growthfactor，VEGF）表达水平明显高于MIA-PaCa2细胞。结论 MIA-PaCa2-3细胞尾静脉注射可建立高肝转移率的裸鼠胰腺癌模型，借助荧光示踪可以简便、可靠地观察肝脏转移肿瘤的生长情况，适于胰腺癌肝转移机制研究和抗肝转移药物的筛选。

Objective To establish a pancreatic cancer nude mice model with high metastatic potential for study of the mechanism and drug screening of pancreatic cancer liver metastasis. Methods Human pancreatic cancer cell lines MIA-PaCa2 were injected into the spleen, and cancer cells from liver metastatic foli of mice were cultured in vitro and subsequently injected into the spleen. After the procedures were performed three times, a sub-line of MIA-PaCa2 with high liver metastatic potential MIA-PaCa2-3 labeled with CM-DiI were injected through the tail vein. Four metastatic related gene expressions including MMP-2, MMP-9, VEGF and bFGF were compared between MIA-PaCa2 and MIA-PaCa2-3 cells by Western-blot. Results Pancreatic cancer models with MIA- PaCa2-3 had a significantly higher metastatic rate （8/10） than its parental cells. The expressions of MMP-2, MMP-9 and VEGF were higher in MIA-PaCa2-3 than in MIA-PaCa2. Conclusions MIA-PaCa2-3 cells demonstrate high metastatic potentials in the liver of nude mice. This model is suitable for study of the mechanism and drug testing for pancreatic cancer metastasis.