高侵袭性膀胱癌细胞系筛选及其裸鼠多器官转移模型的建立  

Screening of a highly invasive bladder cancer cell line and establishment of multiple organ metastatic mouse model

作　　者：徐伟东[1] 许传亮[1] 任善成[1] 王富博[1] 吴承耀[1] 韦荣超[1] 孙颖浩[1] 

机构地区：[1]上海长海医院泌尿外科,上海200433

出　　处：《上海医学》2013年第7期596-599,I0001,共5页Shanghai Medical Journal

摘　　要：目的通过体内培养筛选出高恶性度、高转移潜能的膀胱癌细胞系,构建膀胱癌裸鼠多器官转移模型,以期为进一步转移性膀胱癌的研究和诊疗奠定基础。方法将膀胱癌T24细胞系原位种植于裸鼠膀胱成瘤后（5周）,取出原位瘤进行细胞培养,经胶原酶消化处理后以RPMI1640培养液＋10%胎牛血清培养。采用差异性胰酶消化法进行传代以去除间质细胞。细胞系稳定传代（20代）后原位种植于6只裸鼠膀胱。5周后处死裸鼠,观察肿瘤局部浸润和远处转移情况。取下原位瘤体后,采用免疫组织化学法鉴定是否为膀胱尿路上皮来源。结果成功筛选出高侵袭性膀胱癌细胞系（T24M细胞系）,T24M的细胞形态稳定,已经传至第86代,细胞形态和生长速度均无明显变化,且T24M细胞冻存复苏后的生长状态良好。将T24M细胞系稳定传代后（20代）种植于6只雌性裸鼠膀胱,35d后处死,解剖后发现,6只裸鼠均荷瘤成功且发生远处转移,平均每只裸鼠发生远处转移的器官数为7个（5~9个）,包括髂血管淋巴结、腹膜后淋巴结和肝转移等,转移发生率为6/6,成功建立膀胱癌裸鼠多器官动物模型。免疫组织化学法检测,T24细胞系和T24M细胞系裸鼠膀胱原位肿瘤均高表达角蛋白CKAE1,可证实裸鼠肿瘤均来源于尿路上皮。结论膀胱癌裸鼠多器官转移模型成功建立。T24M细胞系和裸鼠多器官转移模型可用来进一步研究膀胱癌的发生、发展和转移机制。Objective To establish a highly malignant and metastatic bladder cancer cell line and a multiple organ metastatic model in nude mouse,so as to lay the foundations for research on metastatic bladder cancer.Methods Bladder cancer cell line T24was inoculated orthotopically into the bladder of 6nude mice.Five weeks later,the mice were executed and local invasion tumors were harvested.The tissues were digested with collagenase and then cultured with RPMI1640supplemented with 10%fetal bovine serum.Differential tripsin digestion method was used to remove stromal cells.After 20passages,cells were inoculated orthotopically into the bladder of 6nude mice.Five weeks later,the mice were executed and local invasion and distant metastasis were evaluated.The tumors were then harvested for immunohistochemical study.Results A highly invasive bladder cancer cell line（T24M）was successfully established.The cellular morphology remained unchanged after 86passages and the cells could recover effectively after freezing.These T24Mcells were then implanted into the bladders of 6male mice after they were cultured in vitrofor 20passages.The mice were sacrificed 35dlater,and distant metastases were found in the iliac lymph node,the retroperitoneal lymph node and the liver in each mouse.The incidence of tumor metastasis was 6/6and the number of metastasis organs（tumors）was 7（range,5-9）.Nude mice multiple organ metastasis model was successfully established.Immunohistochemical test demonstrated high CKAE1 expression in the T24cells and T24Mcells and the tumors originated from urothelium.Conclusion T24M cell line and bladder cancer multiple organ metastases model have been successfully established.Furthermore,T24Mcells may also be used for the studies on the development,progression and metastasis of bladder cancer.

关 键 词：膀胱癌 原代细胞系 原位模型 T24 

分 类 号：R737.14[医药卫生—肿瘤]