稳定表达人IDO基因的小鼠肺癌原位移植瘤模型的建立

Establishment of an orthotopic transplant tumor model of murine lung cancer with stable expression of human IDO gene in mice

目的 :建立稳定表达人吲哚胺-2,3-双加氧酶(indoleamine-2,3-dioxygenase,IDO)基因的小鼠肺癌原位移植瘤模型。方法 :将稳定表达IDO的小鼠肺癌2LL-EGFP-IDO细胞分别直接种植于小鼠肺脏和皮下,观察2组小鼠的生存期。通过病理学方法检测肺部病灶的成瘤情况。应用0.9%氯化钠溶液(模型组)、肺积方、IDO抑制剂右旋1-甲基色氨酸和紫杉醇分别干预肺癌原位移植瘤模型小鼠,活体成像法检测肺肿瘤生长的动态变化,蛋白质印迹法检测肺癌原位移植瘤组织中IDO蛋白的表达。结果 :肺癌原位移植瘤模型组小鼠的生存期明显短于皮下移植瘤组(P=0.002)。病理检测结果证实,小鼠肺部存在肿瘤浸润病灶。与模型组比较,肺积方、右旋1-甲基色氨酸和紫杉醇均可抑制小鼠肺癌原位移植瘤的生长(P值均<0.05);模型组肺癌组织中IDO的表达水平明显高于高剂量肺积方组(100 mg·g-1·d-1)、低剂量肺积方组(50 mg·g-1·d-1)和右旋1-甲基色氨酸组(P值均<0.05)。结论 :成功建立2LL-EGFP-IDO细胞小鼠肺癌原位移植瘤模型,且肺积方和右旋1-甲基色氨酸均可抑制小鼠肺癌原位移植瘤的生长及IDO的表达。

Objective：To establish an orthotopic transplant tumor model of murine lung cancer with stable expression of human indoleamine-2,3-dioxygenase（IDO） gene in mice.Methods：The murine lung cancer 2LL-ECFP-IDO cells with stable expression of IDO gene were transplanted into the lung or subcutaneous tissue of mice.The survival time of mice was observed.The tumorigenesis of murine lung cancer cells in lung was detected by pathological examination.The orthotopic transplant tumor model of lung cancer in mice was treated with0.9%NaCI（as a model group）,Chinese medicinal formula Feiji Recipe,IDO inhibitor 1-methyl tryptophan（1-MT） or paclitaxel,respectively.The dynamic change of growth of lung cancer was observed by in vivo optical imaging.The expression level of IDO protein in the orthotopic transplant tumor of murine lung cancer was detected by Western blotting.Results：The survival time of mice-bearing orthotopic transplant tumor of murine lung cancer was shorterned as compared with that of the mice-bearing subcutaneous transplant tumor（P = 0.002）.The pathologic result showed that tumorigenesis could be observed in the lung in mice.As compared with the model group,the growth of orthotopic transplant tumor of lung cancer was inhibited by Feiji Recipe,1-MT and paclitaxel（all P 0.05）.The expression level of IDO protein in the model group was higher than those in high-dose（100 mg-g^（-1）·d^（-1）） Feiji Recipe,low-dose（50 mg·g^（-1）·d^（-1）） Feiji Recipe and 1-MT groups（all P 0.05）.Conclusion：The orthotopic transplant tumor model of murine lung cancer 2LL-EGFP-IDO cells in mice can be successfully established,and the growth of transplant tumor and the IDO expression level can be inhibited by Feiji Recipe and 1-MT.