[Effect of mouse uroplakin II promoter on human bladder cancer cell line].
OBJECTIVE
To study the effect of gene expression of mouse uroplakin II (UPII) promoter on human bladder cell cancer cell line.


METHODS
The mRNA expression of different cell lines was quantified by RT-PCR. Green fluorescent protein (GFP) and luciferase (Luc) were used as reporter genes. The plasmids carrying UPII or GFP were constructed and transfected into human cell lines of bladder transitional cell cancer (BIU-87), kindey cancer (GRC-1), vascular endothelium (EC), lung cancer cell line (A549) and skin fibroblast cell line (Hs27). GFP activity of cells was detected by confocual microscopy and flow cytometry (FCM). Luciferase value was measured by luminometer (microplate) and luciferase to beta-galactosidase ratios (L/G values) were used for evaluating transfection efficiency.


RESULTS
RT-PCR showed high expression level of UPII mRNA in bladder cancer cell line BIU-87, whereas low level or no expression in nonbladder cancer cell lines. The activity of GFP in bladder cancer (BIU-87) cell was higher than that in the other cell lines (5 - 10/HP versus 0 - 2/HP), with 4.34% positive cells in BIU-87 detected by FCM, but no positive cell was found in the other cell lines. L/G values indicated that the luciferase expression in human bladder cancer cells transfected with mouse UPII promoter was 1.8 - 8.2-fold as high as that in the nonbladder cell lines.


CONCLUSIONS
Mouse UPII promoter gene can be expressed in a tissue-specific fashion in human urinary bladder cancer. It is capable of initiating transcription of reporter genes in human bladder cancer cell line.
Zhu
Hong-jian
HJ
Institute of Urology, Peking University, Beijing 100034, China.
Zhang
Zhi-qing
ZQ
Zeng
Xiang-fu
XF
Wei
Shou-shun
SS
Xu
Chun-xiao
CX
Huang
Guo-jin
GJ
Guo
Ying-lu
YL
D000818:Animals; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000628:therapy; D060170:Uroplakin II
D008164:Luminescent Proteins; D008565:Membrane Proteins; C554658:UPK2 protein, human; C554659:Upk2 protein, mouse; D060170:Uroplakin II; D049452:Green Fluorescent Proteins