Detection of methylated apoptosis-associated genes in urine sediments of bladder cancer patients

Martin G. Friedrich, Daniel J. Weisenberger, Jonathan C. Cheng, Shahin Chandrasoma, Kimberly D. Siegmund, Mark L Gonzalgo, Marieta I. Toma, Hartwig Huland, Christine Yoo, Yvonne C. Tsai, Peter W. Nichols, Bernard H. Bochner, Peter A. Jones, Gangning Liang

Research output: Contribution to journalArticle

173 Citations (Scopus)

Abstract

Purpose: There is increasing evidence for a fundamental role for epigenetic silencing of apoptotic pathways in cancer. Changes in DNA methylation can be detected with a high degree of sensitivity, so we used the MethyLight assay to determine how methylation patterns of apoptosis-associated genes change during bladder carcinogenesis and whether DNA methylation could be detected in urine sediments. Experimental Design: We analyzed the methylation status of the 5′ regions of 12 apoptosis-associated genes (ARF, FADD, TNFRSF21, BAX, LITAF, DAPK, TMS-1, BCL2, RASSF1A, TERT, TNFRSF25, and EDNRB) in 18 bladder cancer cell lines, 127 bladder cancer samples, and 37 samples of adjacent normal bladder mucosa using the quantitative MethyLight assay. We also analyzed the methylation status in urine sediments of 20 cancer-free volunteers and 37 bladder cancer patients. Results: The 5′ regions of DAPK, BCL2, TERT, RASSF1A, and TNFRSF25 showed significant increases in methylation levels when compared with nonmalignant adjacent tissue (P ≤ 0.01). Methylation levels of BCL2 were significantly associated with tumor staging and grading (P ≤ 0.01), whereas methylation levels of RASSF1A and ARF were only associated with tumor stage (P ≤ 0.04), and TERT methylation and EDNRB methylation were predictors of tumor grade (P ≤ 0.02). To investigate clinical usefulness for non-invasive bladder cancer detection, we further analyzed the methylation status of the markers in urine samples of patients with bladder cancer. Methylation of DAPK, BCL2, and TERT in urine sediment DNA from bladder cancer patients was detected in the majority of samples (78%), whereas they were unmethylated in the urine sediment DNA from age-matched cancer-free individuals. Conclusions: Our results indicate that methylation of the 5′ region of apoptosis-asscciated genes is a common finding in patients with bladder carcinoma. The ability to detect methylation not only in bladder tissue, but also in urine sediments, suggests that methylation markers are promising tools for noninvasive detection of bladder cancers. Our results also indicate that some methylation markers, such as those in regions of RASSF1A and TNFRSF25, might be of limited ose for detection because they are also methylated in normal bladder tissues.

Original languageEnglish (US)
Pages (from-to)7457-7465
Number of pages9
JournalClinical Cancer Research
Volume10
Issue number22
DOIs
StatePublished - Dec 15 2004
Externally publishedYes

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Urinary Bladder Neoplasms
Methylation
Urine
Apoptosis
Genes
Urinary Bladder
DNA Methylation
Neoplasms
Neoplasm Grading
Neoplasm Staging
DNA
Epigenomics
Volunteers
Carcinogenesis
Mucous Membrane
Research Design

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Friedrich, M. G., Weisenberger, D. J., Cheng, J. C., Chandrasoma, S., Siegmund, K. D., Gonzalgo, M. L., ... Liang, G. (2004). Detection of methylated apoptosis-associated genes in urine sediments of bladder cancer patients. Clinical Cancer Research, 10(22), 7457-7465. https://doi.org/10.1158/1078-0432.CCR-04-0930

Detection of methylated apoptosis-associated genes in urine sediments of bladder cancer patients. / Friedrich, Martin G.; Weisenberger, Daniel J.; Cheng, Jonathan C.; Chandrasoma, Shahin; Siegmund, Kimberly D.; Gonzalgo, Mark L; Toma, Marieta I.; Huland, Hartwig; Yoo, Christine; Tsai, Yvonne C.; Nichols, Peter W.; Bochner, Bernard H.; Jones, Peter A.; Liang, Gangning.

In: Clinical Cancer Research, Vol. 10, No. 22, 15.12.2004, p. 7457-7465.

Research output: Contribution to journalArticle

Friedrich, MG, Weisenberger, DJ, Cheng, JC, Chandrasoma, S, Siegmund, KD, Gonzalgo, ML, Toma, MI, Huland, H, Yoo, C, Tsai, YC, Nichols, PW, Bochner, BH, Jones, PA & Liang, G 2004, 'Detection of methylated apoptosis-associated genes in urine sediments of bladder cancer patients', Clinical Cancer Research, vol. 10, no. 22, pp. 7457-7465. https://doi.org/10.1158/1078-0432.CCR-04-0930
Friedrich, Martin G. ; Weisenberger, Daniel J. ; Cheng, Jonathan C. ; Chandrasoma, Shahin ; Siegmund, Kimberly D. ; Gonzalgo, Mark L ; Toma, Marieta I. ; Huland, Hartwig ; Yoo, Christine ; Tsai, Yvonne C. ; Nichols, Peter W. ; Bochner, Bernard H. ; Jones, Peter A. ; Liang, Gangning. / Detection of methylated apoptosis-associated genes in urine sediments of bladder cancer patients. In: Clinical Cancer Research. 2004 ; Vol. 10, No. 22. pp. 7457-7465.
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abstract = "Purpose: There is increasing evidence for a fundamental role for epigenetic silencing of apoptotic pathways in cancer. Changes in DNA methylation can be detected with a high degree of sensitivity, so we used the MethyLight assay to determine how methylation patterns of apoptosis-associated genes change during bladder carcinogenesis and whether DNA methylation could be detected in urine sediments. Experimental Design: We analyzed the methylation status of the 5′ regions of 12 apoptosis-associated genes (ARF, FADD, TNFRSF21, BAX, LITAF, DAPK, TMS-1, BCL2, RASSF1A, TERT, TNFRSF25, and EDNRB) in 18 bladder cancer cell lines, 127 bladder cancer samples, and 37 samples of adjacent normal bladder mucosa using the quantitative MethyLight assay. We also analyzed the methylation status in urine sediments of 20 cancer-free volunteers and 37 bladder cancer patients. Results: The 5′ regions of DAPK, BCL2, TERT, RASSF1A, and TNFRSF25 showed significant increases in methylation levels when compared with nonmalignant adjacent tissue (P ≤ 0.01). Methylation levels of BCL2 were significantly associated with tumor staging and grading (P ≤ 0.01), whereas methylation levels of RASSF1A and ARF were only associated with tumor stage (P ≤ 0.04), and TERT methylation and EDNRB methylation were predictors of tumor grade (P ≤ 0.02). To investigate clinical usefulness for non-invasive bladder cancer detection, we further analyzed the methylation status of the markers in urine samples of patients with bladder cancer. Methylation of DAPK, BCL2, and TERT in urine sediment DNA from bladder cancer patients was detected in the majority of samples (78{\%}), whereas they were unmethylated in the urine sediment DNA from age-matched cancer-free individuals. Conclusions: Our results indicate that methylation of the 5′ region of apoptosis-asscciated genes is a common finding in patients with bladder carcinoma. The ability to detect methylation not only in bladder tissue, but also in urine sediments, suggests that methylation markers are promising tools for noninvasive detection of bladder cancers. Our results also indicate that some methylation markers, such as those in regions of RASSF1A and TNFRSF25, might be of limited ose for detection because they are also methylated in normal bladder tissues.",
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T1 - Detection of methylated apoptosis-associated genes in urine sediments of bladder cancer patients

AU - Friedrich, Martin G.

AU - Weisenberger, Daniel J.

AU - Cheng, Jonathan C.

AU - Chandrasoma, Shahin

AU - Siegmund, Kimberly D.

AU - Gonzalgo, Mark L

AU - Toma, Marieta I.

AU - Huland, Hartwig

AU - Yoo, Christine

AU - Tsai, Yvonne C.

AU - Nichols, Peter W.

AU - Bochner, Bernard H.

AU - Jones, Peter A.

AU - Liang, Gangning

PY - 2004/12/15

Y1 - 2004/12/15

N2 - Purpose: There is increasing evidence for a fundamental role for epigenetic silencing of apoptotic pathways in cancer. Changes in DNA methylation can be detected with a high degree of sensitivity, so we used the MethyLight assay to determine how methylation patterns of apoptosis-associated genes change during bladder carcinogenesis and whether DNA methylation could be detected in urine sediments. Experimental Design: We analyzed the methylation status of the 5′ regions of 12 apoptosis-associated genes (ARF, FADD, TNFRSF21, BAX, LITAF, DAPK, TMS-1, BCL2, RASSF1A, TERT, TNFRSF25, and EDNRB) in 18 bladder cancer cell lines, 127 bladder cancer samples, and 37 samples of adjacent normal bladder mucosa using the quantitative MethyLight assay. We also analyzed the methylation status in urine sediments of 20 cancer-free volunteers and 37 bladder cancer patients. Results: The 5′ regions of DAPK, BCL2, TERT, RASSF1A, and TNFRSF25 showed significant increases in methylation levels when compared with nonmalignant adjacent tissue (P ≤ 0.01). Methylation levels of BCL2 were significantly associated with tumor staging and grading (P ≤ 0.01), whereas methylation levels of RASSF1A and ARF were only associated with tumor stage (P ≤ 0.04), and TERT methylation and EDNRB methylation were predictors of tumor grade (P ≤ 0.02). To investigate clinical usefulness for non-invasive bladder cancer detection, we further analyzed the methylation status of the markers in urine samples of patients with bladder cancer. Methylation of DAPK, BCL2, and TERT in urine sediment DNA from bladder cancer patients was detected in the majority of samples (78%), whereas they were unmethylated in the urine sediment DNA from age-matched cancer-free individuals. Conclusions: Our results indicate that methylation of the 5′ region of apoptosis-asscciated genes is a common finding in patients with bladder carcinoma. The ability to detect methylation not only in bladder tissue, but also in urine sediments, suggests that methylation markers are promising tools for noninvasive detection of bladder cancers. Our results also indicate that some methylation markers, such as those in regions of RASSF1A and TNFRSF25, might be of limited ose for detection because they are also methylated in normal bladder tissues.

AB - Purpose: There is increasing evidence for a fundamental role for epigenetic silencing of apoptotic pathways in cancer. Changes in DNA methylation can be detected with a high degree of sensitivity, so we used the MethyLight assay to determine how methylation patterns of apoptosis-associated genes change during bladder carcinogenesis and whether DNA methylation could be detected in urine sediments. Experimental Design: We analyzed the methylation status of the 5′ regions of 12 apoptosis-associated genes (ARF, FADD, TNFRSF21, BAX, LITAF, DAPK, TMS-1, BCL2, RASSF1A, TERT, TNFRSF25, and EDNRB) in 18 bladder cancer cell lines, 127 bladder cancer samples, and 37 samples of adjacent normal bladder mucosa using the quantitative MethyLight assay. We also analyzed the methylation status in urine sediments of 20 cancer-free volunteers and 37 bladder cancer patients. Results: The 5′ regions of DAPK, BCL2, TERT, RASSF1A, and TNFRSF25 showed significant increases in methylation levels when compared with nonmalignant adjacent tissue (P ≤ 0.01). Methylation levels of BCL2 were significantly associated with tumor staging and grading (P ≤ 0.01), whereas methylation levels of RASSF1A and ARF were only associated with tumor stage (P ≤ 0.04), and TERT methylation and EDNRB methylation were predictors of tumor grade (P ≤ 0.02). To investigate clinical usefulness for non-invasive bladder cancer detection, we further analyzed the methylation status of the markers in urine samples of patients with bladder cancer. Methylation of DAPK, BCL2, and TERT in urine sediment DNA from bladder cancer patients was detected in the majority of samples (78%), whereas they were unmethylated in the urine sediment DNA from age-matched cancer-free individuals. Conclusions: Our results indicate that methylation of the 5′ region of apoptosis-asscciated genes is a common finding in patients with bladder carcinoma. The ability to detect methylation not only in bladder tissue, but also in urine sediments, suggests that methylation markers are promising tools for noninvasive detection of bladder cancers. Our results also indicate that some methylation markers, such as those in regions of RASSF1A and TNFRSF25, might be of limited ose for detection because they are also methylated in normal bladder tissues.

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