Tanaka Motoysohi, Gee Jason R, De La Cerda Jorge, Rosser Charles J, Zhou Jain-Hua, Benedict William F, Grossman H Barton
Department of Urology, University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030, USA.
J Urol. 2003 Sep;170(3):975-8. doi: 10.1097/01.ju.0000073209.65128.c1.
Orthotopic models of bladder cancer mimic the normal microenvironment and provide an opportunity to study new therapies for superficial bladder cancer. The use of green fluorescent protein (GFP) transduced cells provides a sensitive way of monitoring this disease. We investigated whether examining voided urine for GFP expressing cells would indicate the presence of GFP producing tumors in an orthotopic bladder tumor model in nude mice.
The human bladder cancer cell lines KU-7, UM-UC-3 and UM-UC-14 were used. GFP transductants were generated after transfection with pEGFP-N3, followed by G418 selection. After the cells were inoculated in an orthotopic model of superficial bladder cancer voided urine was collected on slides weekly for 3 weeks and observed for GFP expressing cells by fluorescence microscopy. Bladder tumor imaging for GFP was performed in surgically exposed bladders to determine the tumor incidence.
KU-7 GFP cells produced tumors in all 16 mice on whole bladder GFP imaging. UM-UC-3 and UM-UC-14 GFP cells produced tumors in 8 of 12 (67%) and 18 of 25 (72%) mice, respectively. The rate of GFP positive cells in spontaneously voided urine varied by cell line and increased with time but it was generally less than the rate of detection by whole bladder GFP imaging. All mice with GFP expressing cells in the urine had GFP expressing bladder tumors.
Examining urine for GFP expressing cells is less sensitive than imaging surgically exposed bladders but it is 100% specific.
膀胱癌原位模型可模拟正常微环境，为研究浅表性膀胱癌的新疗法提供了契机。使用绿色荧光蛋白（GFP）转导细胞为监测这种疾病提供了一种灵敏的方法。我们研究了检测晨尿中表达GFP的细胞是否能表明在裸鼠原位膀胱肿瘤模型中存在产生GFP的肿瘤。
使用人膀胱癌细胞系KU-7、UM-UC-3和UM-UC-14。用pEGFP-N3转染后产生GFP转导子，随后进行G418筛选。将细胞接种到浅表性膀胱癌原位模型后，每周在载玻片上收集晨尿，持续3周，并通过荧光显微镜观察表达GFP的细胞。对手术暴露的膀胱进行GFP膀胱肿瘤成像以确定肿瘤发生率。
在全膀胱GFP成像中，KU-7 GFP细胞在所有16只小鼠中均产生肿瘤。UM-UC-3和UM-UC-14 GFP细胞分别在12只小鼠中的8只（67%）和25只小鼠中的18只（72%）产生肿瘤。晨尿中GFP阳性细胞的比例因细胞系而异，并随时间增加，但通常低于全膀胱GFP成像的检测率。所有尿液中含有表达GFP细胞的小鼠均有表达GFP的膀胱肿瘤。
检测尿液中表达GFP的细胞比成像手术暴露的膀胱敏感性低，但特异性为100%。
