人绒毛膜癌鼠肺转移模型的建立及其生物学特性的初步观察

目的 建立1种较为符合临床特征的人绒毛膜癌(绒癌)肺转移动物模型,初步观察其生物学特性,并探索成功建立模型的合适的细胞浓度、细胞数量.方法 将40只5～6周龄重症联合免疫缺陷(SCID)小鼠随机分为4组,每组各10只,人绒癌细胞系JEG-3分别以1×107个细胞/ml×0.1 ml(A组)、5×106个细胞/ml×0.2 ml(B组)、1×106个细胞/ml×0.1 ml(C组)注入SCID小鼠尾静脉,另设对照组.接种后每3天观察1次小鼠状态,称量1次小鼠的体质量,绘制小鼠体质量变化曲线;当小鼠处于濒死状态时,用小动物成像CT(Micro CT)系统检查肺部有无转移灶、转移灶数目及大小.Micro CT检查后,解剖SCID小鼠,取肺部转移灶进行细胞的原代培养并行病理取材,HE染色、病理检查,免疫组织化学方法检测人绒毛膜促性腺激素β亚单位(β-hCG)的表达情况;原代培养细胞行免疫组织化学检测β-hCG的表达情况,并行染色体核型分析.结果 A组小鼠接种时全部死亡;C组接种时死亡2只,存活的8只在(30.0±2.0)d时,处于濒死状态,进行解剖未发现肺转移灶.B组接种时死亡3只,存活的7只在(18.0±2.0)d时,处于濒死状态;Micro CT检查,肺转移率为5/7,肺转移灶直径1.5～3.5 mm;病理检查证实为绒癌.原代培养的细胞行人绒癌标志物β-hOG染色,呈阳性表达;原代培养的细胞染色体核型分析为单体、三体和多体人源性染色体,细胞染色体数目19～128条,染色体众数为70～79条,符合恶性肿瘤的遗传学特征.结论 通过尾静脉注射JEG-3细胞可以成功建立人绒癌SCID小鼠肺转移模型,5×106个细胞/ml×0.2 ml为建立模型的较为合适的细胞浓度及容积.

Lung metastasis of human choriocarcinoma in mice: establishment of experimental metastatic model and its biological characteristics

Objective To establish a satisfactory lung metastasis model of human choriocarcinoma using severe combined immunedeficient(SCID)mice and explore the appropriate cell concentration for the model.Methods Forty SCID mice aged between 5-6 weeks were randomly difided into four groups.1×107 cells/ml ×0.1 ml.5×106 cells/ml×0.2 ml and 1×106 cells/ml×0.1 ml of human choriocarcinoma cells JEG-3 were respectively injected in SCID mice of experimental groups by lateral tail vein,the remain group was assigned to the control group.The status and weisht of mice were observed every three days.When these mice were being dying.the size and the number of the lesions of lung metastasis in every mouse were inspected with Micro CT.After Micro CT inspection,the SCID mice were executed dissected to note whether there were tumors on all organ surfaces witll naked eyes.then made pathological sections from the metastaticfoci of fresh lung tissues,and cultured primarily cells and purified cells and passaged cells isolated from the same metastastic foci.The pathological sections were observed under the microscope.The special antigen human chorionic gonadotropin-beta subunit(β-hCG)of the choriocareinoma cells was immunohistochemically detected in the pathological sections and the cells out of cultured primarily cells.The chromosomes of the cells out of cultured primarily cells were analysed.Results Of the group inoeutated 1×107 cells/ml×0.1 ml.all mice died when inoculating.In the group of 5×106 cells/ml×0.2 ml,when inoculating, 3 mice died; the remain 7 mice were being dying on ( 18. 0 ±2. 0) days after injection. 5 of them, there were 1 - 3 lesions of lung metastasis after Micro CT inspection in each mice, and the diameter of the tumors lesions reached 1.5 - 3.5 ram, which was choriocarcinoma confirmed by pathological sections.The special antigen β-hCG was detected by immunohistoehemical method in the pathological sections of pulmonary tissue with tumor and in the cells, which were purified and passaged from being cultured primarily cells isolated from metastastic foci of fresh lung tissues from the SCID mice. The chromosome numbers of these cells out of cultured primarily ceils were variety from 19 to 128, and medal numbers were variety from 70 to 79. Conclusions We successfully established the lung metastatic model of human choriocarcinoma in SCID mice by injecting JEG-3 cells into lateral tail vein, of which 5 × 106 cells/ml × 0. 2 ml is the suitable concentration and volume for the model.