Restoration of expression of transforming growth factor-β Type II receptor in murine renal cell carcinoma (renca) cells by 5-Aza-2′- deoxycytidine

Qiang Zhang; Jonathan N. Rubenstein; Victoria C. Liu; Irwin Park; Thomas Jang; Chung Lee

doi:10.1016/j.lfs.2004.10.021

Restoration of expression of transforming growth factor-β Type II receptor in murine renal cell carcinoma (renca) cells by 5-Aza-2′- deoxycytidine

Qiang Zhang, Jonathan N. Rubenstein, Victoria C. Liu, Irwin Park, Thomas Jang, Chung Lee

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

The murine renal cell carcinoma (Renca) cells are insensitive to TGF-β due to a lack of TGF-β type II receptor (TβR-II). The objective of the present study is to determine the mechanism of this loss of sensitivity to TGF-β in Renca cells. Renca cells were cultured and treated with 5-Aza-2′-Deoxycytidine (5-Aza), a specific inhibitor of methylation. Expression of TGF-β type I receptor (TβRI) and TβRII was determined by RT-PCR and Western blot analysis before and after the treatment of Renca cells with 5-Aza. The expression of phosphorylated Smad2 (P-Smad2) was determined by Western blot analysis. TGF-β levels in the conditioned medium were measured by ELISA. Renca cells did not express TβR-II prior to 5-Aza treatment. After 5-Aza treatment, these cells expressed TβR-II at both mRNA and protein levels, which corresponded to the restoration of sensitivity to TGF-β by an increase in P-Smad2. Levels of TGF-β1 were similar before and after 5-Aza treatment. Results of the present study indicated that, in Renca cells, the loss of sensitivity to TGF-β is likely due to a promoter hypermethylation in the TβR-II gene.

Original language	English (US)
Pages (from-to)	1159-1166
Number of pages	8
Journal	Life Sciences
Volume	76
Issue number	10
DOIs	https://doi.org/10.1016/j.lfs.2004.10.021
State	Published - Jan 21 2005

Funding

This work was supported in part, by grants from National Cancer Institute (CA 60553, CA 90386), the American Fundation For Urologic Disease For Post-doctor Fellow (QZ), and Riba Urology Fellowship of Northwestern University (JNR).

Keywords

Promoter methylation
Renal cell carcinoma
TGF-β Signaling
TGF-β receptors

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
General Pharmacology, Toxicology and Pharmaceutics

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10.1016/j.lfs.2004.10.021

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title = "Restoration of expression of transforming growth factor-β Type II receptor in murine renal cell carcinoma (renca) cells by 5-Aza-2′- deoxycytidine",

abstract = "The murine renal cell carcinoma (Renca) cells are insensitive to TGF-β due to a lack of TGF-β type II receptor (TβR-II). The objective of the present study is to determine the mechanism of this loss of sensitivity to TGF-β in Renca cells. Renca cells were cultured and treated with 5-Aza-2′-Deoxycytidine (5-Aza), a specific inhibitor of methylation. Expression of TGF-β type I receptor (TβRI) and TβRII was determined by RT-PCR and Western blot analysis before and after the treatment of Renca cells with 5-Aza. The expression of phosphorylated Smad2 (P-Smad2) was determined by Western blot analysis. TGF-β levels in the conditioned medium were measured by ELISA. Renca cells did not express TβR-II prior to 5-Aza treatment. After 5-Aza treatment, these cells expressed TβR-II at both mRNA and protein levels, which corresponded to the restoration of sensitivity to TGF-β by an increase in P-Smad2. Levels of TGF-β1 were similar before and after 5-Aza treatment. Results of the present study indicated that, in Renca cells, the loss of sensitivity to TGF-β is likely due to a promoter hypermethylation in the TβR-II gene.",

keywords = "Promoter methylation, Renal cell carcinoma, TGF-β Signaling, TGF-β receptors",

author = "Qiang Zhang and Rubenstein, {Jonathan N.} and Liu, {Victoria C.} and Irwin Park and Thomas Jang and Chung Lee",

note = "Funding Information: This work was supported in part, by grants from National Cancer Institute (CA 60553, CA 90386), the American Fundation For Urologic Disease For Post-doctor Fellow (QZ), and Riba Urology Fellowship of Northwestern University (JNR).",

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doi = "10.1016/j.lfs.2004.10.021",

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AU - Zhang, Qiang

AU - Rubenstein, Jonathan N.

AU - Liu, Victoria C.

AU - Park, Irwin

AU - Jang, Thomas

AU - Lee, Chung

N1 - Funding Information: This work was supported in part, by grants from National Cancer Institute (CA 60553, CA 90386), the American Fundation For Urologic Disease For Post-doctor Fellow (QZ), and Riba Urology Fellowship of Northwestern University (JNR).

PY - 2005/1/21

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N2 - The murine renal cell carcinoma (Renca) cells are insensitive to TGF-β due to a lack of TGF-β type II receptor (TβR-II). The objective of the present study is to determine the mechanism of this loss of sensitivity to TGF-β in Renca cells. Renca cells were cultured and treated with 5-Aza-2′-Deoxycytidine (5-Aza), a specific inhibitor of methylation. Expression of TGF-β type I receptor (TβRI) and TβRII was determined by RT-PCR and Western blot analysis before and after the treatment of Renca cells with 5-Aza. The expression of phosphorylated Smad2 (P-Smad2) was determined by Western blot analysis. TGF-β levels in the conditioned medium were measured by ELISA. Renca cells did not express TβR-II prior to 5-Aza treatment. After 5-Aza treatment, these cells expressed TβR-II at both mRNA and protein levels, which corresponded to the restoration of sensitivity to TGF-β by an increase in P-Smad2. Levels of TGF-β1 were similar before and after 5-Aza treatment. Results of the present study indicated that, in Renca cells, the loss of sensitivity to TGF-β is likely due to a promoter hypermethylation in the TβR-II gene.

AB - The murine renal cell carcinoma (Renca) cells are insensitive to TGF-β due to a lack of TGF-β type II receptor (TβR-II). The objective of the present study is to determine the mechanism of this loss of sensitivity to TGF-β in Renca cells. Renca cells were cultured and treated with 5-Aza-2′-Deoxycytidine (5-Aza), a specific inhibitor of methylation. Expression of TGF-β type I receptor (TβRI) and TβRII was determined by RT-PCR and Western blot analysis before and after the treatment of Renca cells with 5-Aza. The expression of phosphorylated Smad2 (P-Smad2) was determined by Western blot analysis. TGF-β levels in the conditioned medium were measured by ELISA. Renca cells did not express TβR-II prior to 5-Aza treatment. After 5-Aza treatment, these cells expressed TβR-II at both mRNA and protein levels, which corresponded to the restoration of sensitivity to TGF-β by an increase in P-Smad2. Levels of TGF-β1 were similar before and after 5-Aza treatment. Results of the present study indicated that, in Renca cells, the loss of sensitivity to TGF-β is likely due to a promoter hypermethylation in the TβR-II gene.

KW - Promoter methylation

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KW - TGF-β Signaling

KW - TGF-β receptors

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Restoration of expression of transforming growth factor-β Type II receptor in murine renal cell carcinoma (renca) cells by 5-Aza-2′- deoxycytidine

Abstract

Funding

Keywords

ASJC Scopus subject areas

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