Korean J Urol.  2007 Sep;48(9):915-920. 10.4111/kju.2007.48.9.915.

Analysis of Human V-erbA Related EAR-3 Gene Expression between Transitional Cell Carcinoma and Normal Tissue in Bladder Cancer

• Ham WS
• Lee JH
• Yu HS
• Choi YD

• KMID: 2061291
• DOI: http://doi.org/10.4111/kju.2007.48.9.915

Abstract

PURPOSE
The prognosis of bladder cancer is related to tumor grade and stage. Because these pathological changes are preceded by molecular alterations, new molecular markers are needed in early diagnosis. New target molecular biomarkers can be differentially expressed genes(DEGs) between normal and cancer tissues. We tried to find a new DEG and demonstrated that it may be related to the development of the bladder cancer.
MATERIALS AND METHODS
Cancer tissues were obtained from 39 patients with urothelial cell carcinoma, treated by transurethral resection of tumor (TURB) since 2002. Normal bladder tissues were obtained from the same patients during TURB. We compared the mRNA profiles between normal and cancer tissues using annealing control primer(ACP)-based Genefishing(TM) PCR to identify the DEGs in normal and cancer tissues of one same patient. To validate the result of ACP-based GeneFishing(TM) PCR, reverse transcription-polymerase chain reaction(RT-PCR) was performed on those of 39 patients.
RESULTS
According to the result of ACP-based Genefishing(TM) PCR, EAR-3 gene was only present or markedly upregulated in normal tissue, compared with cancer tissues. The expression pattern that EAR-3 gene was downregulated in cancer tissues, irrespective of the clinicopathologic parameters was confirmed by RT-PCR in 39 patients.
CONCLUSIONS
EAR-3 gene was downregulated in cancer tissues, irrespective of clinicopathologic parameters, compared with normal tissues in the bladder of the same patient. Therefore, we suggested that EAR-3 gene may be also play a role in bladder cancer development.

Keyword

Bladder cancer; ACP-based GeneFishing(TM) PCR; Reverse transcriptase polymerase chain reaction; EAR-3 gene

MeSH Terms

Biological Markers
Carcinoma, Transitional Cell*
Early Diagnosis
Gene Expression*
Humans*
Polymerase Chain Reaction
Prognosis
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger
Urinary Bladder Neoplasms*
Urinary Bladder*
Biological Markers
RNA, Messenger

Figure

• 

  Fig. 1 Results of annealing control primer (ACP)-based polymerase chain reaction (PCR) for identification of differentially expressed genes (DEGs) from normal and cancer tissue. Arrowheads indicate 12 differential cDNA bands, excised from the gel for further cloning and sequencing and 35 band is found to be EAR-3 gene according to the sequence homology search.

• 

  Fig. 2 Reverse transcription-polymerase chain reaction (RT-PCR) product analysis by agarose gel electrophoresis and ethidium bromide staining showing the intensity of EAR-3 gene expression in bladder cancer tissues (C) and normal bladder tissue (N) for each patient. The second line represents internal control β-actin expression.

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