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Decreased expression of REIC/Dkk-3 in human renal clear cell carcinoma
Kyouhei Kurose1, Masakiyo Sakaguchi, Yasutomo Nasu
1Department of Urology, Okayama University Graduate School of Medicine and Dentistry and Faculty of Health Sciences, Okayama, Japan.
Purpose:
We examined the expression of REIC/Dkk-3, a possible candidate for a tumor suppressor gene, in human renal clear cell carcinoma (RCCC) cell lines and sporadic RCCC surgical specimens.
Materials And Methods:
Human RCCC cell lines (Caki-1, Caki-2, ACHN and KPK-1) and several control cell lines were used to examine the expression of REIC/Dkk-3 mRNA and characterize a newly raised antibody specific for REIC/Dkk-3 protein. Pairs of cancerous and adjacent noncancerous tissues were obtained from 20 patients with RCCC. Of them 17 and 7 cases were analyzed by real-time quantitative reverse transcriptase-polymerase chain reaction, and by Western blot analysis and/or immunohistochemical analysis, respectively.
Results:
The decreased expression of REIC/Dkk-3 mRNA and protein in human RCCC cell lines, and the specificity of the new antibody were confirmed. In a real-time quantitative reverse transcriptase-polymerase chain reaction study using 17 pairs of RCCC and adjacent normal tissues REIC/Dkk-3 mRNA levels were significantly decreased in carcinoma tissues (by 25% to approximately 95% in 15 pairs). Western blot analysis and immunohistochemistry revealed a significant decrease in REIC/Dkk-3 protein levels in 6 of the 7 and 13 of the 14 RCCC cases analyzed, respectively.
Conclusions:
The decrease in REIC/Dkk-3 mRNA and protein levels was observed irrespective of tumor grade and stage, indicating the involvement of REIC/Dkk-3 in an initial step of malignant conversion. Consequently REIC/Dkk-3 could be a new molecular target for therapeutic measures against RCCC.
Insights
The expression of REIC/Dkk-3 (Reduced Expression in Immortalized Cells/Dickkopf-3) mRNA and protein is significantly decreased in renal clear cell carcinoma (RCCC). This suggests REIC/Dkk-3 is involved in early cancer development and may be a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Renal clear cell carcinoma (RCCC) is a significant health concern.
- Identifying novel tumor suppressor genes is crucial for understanding RCCC pathogenesis.
- REIC/Dkk-3 (Reduced Expression in Immortalized Cells/Dickkopf-3) is a potential tumor suppressor gene.
Purpose of the Study:
- To investigate the expression levels of REIC/Dkk-3 in RCCC.
- To evaluate REIC/Dkk-3 as a potential tumor suppressor in RCCC.
- To characterize a novel antibody for REIC/Dkk-3 protein detection.
Main Methods:
- Analysis of REIC/Dkk-3 mRNA and protein expression in RCCC cell lines.
- Utilized real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) on 17 RCCC tissue pairs.
- Employed Western blot and immunohistochemical analysis on RCCC surgical specimens.
Main Results:
- REIC/Dkk-3 mRNA levels were significantly reduced in 15 out of 17 RCCC tissue pairs.
- REIC/Dkk-3 protein levels were significantly decreased in RCCC tissues (6/7 by Western blot, 13/14 by immunohistochemistry).
- A novel antibody demonstrated specificity for REIC/Dkk-3 protein.
Conclusions:
- Downregulation of REIC/Dkk-3 occurs early in RCCC development, irrespective of tumor grade and stage.
- REIC/Dkk-3 plays a role in the initial stages of malignant transformation in RCCC.
- REIC/Dkk-3 represents a potential novel molecular target for RCCC therapies.
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