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A potential for tissue restrictive gene therapy in renal cell carcinoma using MN/CA IX promoter
Yen-Chuan Ou1, Thomas A Gardner, Chinghaj Kao
1Division of Urology, Department of Surgery, Taichung Veterans General Hospital, National Yang-Ming University School of Medicine, Taiwan. ycou@vghtc.vghtc.gov.tw
Background:
The expression of the MN/carbonic anhydrase IX (CA IX) protein is detected in renal cell carcinoma (RCC). A tumor-specific MN promoter was designed and evaluated.
Materials And Methods:
Western blot, reverse transcription-PCR and Northern blot for MN expression were performed on Hela cells, LNCaP and four human RCC cell lines. The MN promoter (554 bp) was cloned into pGL3 luciferase reporter vectors and their activities were tested.
Results:
Hela cells, SKRC-31 and SKRC-38 expressed high levels of MN RNA and MN protein. The MN promoter has 5- to 10-fold higher activity in MN-positive cell lines than in MN-negative cell lines. It could enhance the activity of the basic SV40 early promoter (6 to 25 fold) in MN-positive cells but not in MN-negative cells.
Conclusion:
MN expression in RCC may derive from altered expression of transcriptional factor(s), which binds to the MN enhancer element, and can be used for tumor specific toxic gene therapy for RCC.
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