小鼠尿路斑块2启动子对人细胞特异性及表达调控研究  

Effect of mouse UroplakinII promoter on human cells and their transcriptional control

作　　者：朱宏建[1] 张智清[2] 曾祥福[3] 魏守顺[3] 徐春晓[2] 黄国锦[2] 郭应禄[1] 

机构地区：[1]北京大学第一医院北京大学泌尿外科研究所,100034 [2]中国预防医学科学院病毒学研究所 [3]武警总医院泌尿外科

出　　处：《中华实验外科杂志》2003年第11期1016-1018,共3页Chinese Journal of Experimental Surgery

摘　　要：目的　研究小鼠尿路斑块 2 (UPⅡ )启动子的缺失体在人细胞中启动活性。方法　应用脂质体转染技术 ,将含不同长度的小鼠UPⅡ启动子片段的重组质粒转染入人不同组织细胞系 ,利用共聚焦激光扫描显微镜、流式细胞仪和微板检测仪观察报告基因绿色荧光蛋白 (GFP)和荧光素酶 (Luc)的表达情况。结果　UPⅡ在膀胱癌细胞BIU 87表达GFP较非膀胱癌细胞多 ( 4 .3 4%对 0 )。UPⅡ 1、UPⅡ 2、UPⅡ 3在各组织细胞中表达GFP的数目较UPⅡ小 ,UPⅡ 4在各细胞中均无表达。UPⅡ在BIU 87表达Luc的量是其他组织细胞的 1.8～ 8.2倍。UPⅡ 1、UPⅡ 2、UPⅡ 3在各组织细胞中表达Luc的量均减少 ,UPⅡ 4在各组织细胞中几乎无表达。结论　小鼠UPⅡ启动子在人膀胱移行细胞中具有特异性启动活性 ,启动子上游 1.5× 10 3 区域是增强子序列 ,下游 14 3bp区域是核心启动子序列。Objective To study the effect of mouse UroplakinⅡ (UPⅡ) promoter and fragment on human cells.Methods Green fluorescent protein (GFP) and luciferase (Luc) were used as the reporter genes.The plasmids carrying UPⅡ,UPⅡ-1,UPⅡ-2,UPⅡ-3,UPⅡ-4 and GFP or Luc were constructed and transfected into different human cell lines.GFP activity of cells was detected by a confocual laser scan microscope and flow cytometry (FCM).Luc values were measured by luminometer (microplate).Results The activity of UPII promoter in bladder cancer (BIU-87) cells was higher than in other cell lines (4.34 % vs 0).The activity of UPⅡ-1,UPⅡ-2 and UPⅡ-3 in BIU-87 cells was lower than that of UPII.The activity of UPⅡ-4 was hardly detected in all tissue cell lines by FCM.L/G values showed the luciferase expression from mouse UPII promoter in human bladder cancer cells is 1.8-8.2 folds higher than in nonbladder cell lines.Furthermore,the activity of UPⅡ-1,UPⅡ-2 was significantly decreased but was bladder-specific.The activity of 3' flank 143 bp of UPⅡ promoter was detected in both bladder cancer and lung cancer cell lines.Conclusion Mouse UroplakinⅡ promoter is active in human cells and is also tissue-specific to bladder cells.Enhancer is contained in the region of 1.5 kb of 5’ flank of mouse UroplakinⅡ promoter.Core promoter is located in the region of 143 bp of 3’end.

关 键 词：小鼠 尿路斑块2 启动子 人细胞特异性 基因表达 膀胱肿瘤 

分 类 号：R73-3[医药卫生—肿瘤]