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Carcinogenesis Advance Access published online on December 12, 2005
Carcinogenesis, doi:10.1093/carcin/bgi299
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© The Author 2005. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org
Received September 13, 2005
Revised November 8, 2005
Accepted November 27, 2005

CANCER BIOLOGY

Decreased expression of the human stem cell marker, rex-1 (zfp-42), in renal cell carcinoma

Jay D. Raman 1, Nigel P. Mongan 2, Limin Liu 2, Satish K. Tickoo 3, David M. Nanus 4, Douglas S. Scherr 1, and Lorraine J. Gudas 2 *

1 Department of Urology, The New York-Presbyterian Hospital, Weill Medical College of Cornell University, New York, NY
2 Department of Pharmacology, The New York-Presbyterian Hospital, Weill Medical College of Cornell University, New York, NY
3 Department of Pathology, The New York-Presbyterian Hospital, Weill Medical College of Cornell University, New York, NY
4 Division of Hematology and Medical Oncology, The New York-Presbyterian Hospital, Weill Medical College of Cornell University, New York, NY

* To whom correspondence should be addressed.
Lorraine J. Gudas, E-mail: ljgudas{at}med.cornell.edu


  Abstract

The Rex-1 (Zfp-42) gene encodes a zinc finger family transcription factor which is highly expressed in mouse and human embryonic stem (ES) cells. It is one of several gene markers used to identify human stem cells. While several organs are known to harbor adult human stem cells, the presence and distribution of stem cells in both the normal and neoplastic adult kidney remains largely unknown. In this study we evaluated Rex-1 mRNA and protein expression in normal and malignant kidney tissue specimens from human patients. Rex-1 mRNA expression was determined using both reverse transcription and real-time PCR. REX1 protein expression was assessed by Western analysis and immunohistochemistry, using an affinity-purified, polyclonal antibody to the REX1 protein. We found that fourteen of 15 (93%) non-tumor renal parenchymal specimens demonstrated Rex-1 mRNA, compared with 5 of 14 (36%) renal tumors (p<0.005). REX1 protein expression was detected in 21 of 23 (91%) non-tumor and in 7 of 19 (37%) tumor specimens (p<0.001). Furthermore, in 6 of these 7 renal tumor specimens where REX1 protein expression was detected, the levels were at least three-fold lower than those in adjacent, normal kidney tissue. There were no differences in Rex-1 mRNA or protein expression among the various histologic subtypes of renal tumors (clear cell carcinoma, papillary carcinoma, chromophobe carcinoma, and oncocytoma). Immunohistochemical staining confirmed the absence of REX1 in three renal tumor specimens (two clear cell and one papillary carcinoma), while the REX1 protein was detected in a small percentage of proximal tubular cells in normal renal tissue. Immunohistochemical staining of another stem cell marker, OCT4, demonstrated a similar pattern of protein expression in a small percentage of normal renal proximal tubular cells. In summary, we were able to detect Rex-1 mRNA and protein expression in over 90% of normal renal parenchymal specimens, and we observed a significant reduction in REX1 expression in renal tumor specimens of all histologic subtypes.

Keywords: kidney cancer; proximal tubule; kidney stem cell; zfp42; REX1.
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