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Quantitative detection of RASSF1A DNA promoter methylation in tumors and serum of patients with serous epithelial ovarian cancer.

Publication ,  Journal Article
Bondurant, AE; Huang, Z; Whitaker, RS; Simel, LR; Berchuck, A; Murphy, SK
Published in: Gynecol Oncol
December 2011

OBJECTIVE: Detection of cell free tumor-specific DNA methylation has been proposed as a potentially useful noninvasive mechanism to detect malignancies, including ovarian cancer, and to monitor response to treatment. However, there are few easily implemented quantitative approaches available for DNA methylation analysis. Our objectives were to develop an absolute quantitative method for detection of DNA methylation using RASSF1A, a known target of promoter methylation in ovarian cancer, and test the ability to detect RASSF1A methylation in tumors and serum specimens of women with ovarian cancer. METHODS: Bisulfite modified DNAs were subjected to real time PCR using nondiscriminatory PCR primers and a probe with sequence containing a single CpG site, theoretically able to capture the methylation status of that CpG for every allele within a given specimen. Input DNA was normalized to ACTB levels detected simultaneously by assay multiplexing. Methylation levels were established by comparison to results obtained from universally methylated DNA. RESULTS: The assay was able to detect one methylated RASSF1A allele in 100,000 unmethylated alleles. RASSF1A was methylated in 54 of 106 (51%) invasive serous ovarian cancers analyzed and methylation status was concordant in 20/20 matched preoperative serum-tumor pairs. Serial serum specimens taken over the course of treatment for 8 of 9 patients showed fluctuations in RASSF1A methylation concomitant with disease status. CONCLUSIONS: This novel assay provides a real-time PCR-based method for absolute quantitation of DNA methylation. Our results support feasibility of monitoring RASSF1A methylation from serum samples taken over the course of treatment from women with ovarian cancer.

Duke Scholars

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Published In

Gynecol Oncol

DOI

EISSN

1095-6859

Publication Date

December 2011

Volume

123

Issue

3

Start / End Page

581 / 587

Location

United States

Related Subject Headings

  • Tumor Suppressor Proteins
  • Real-Time Polymerase Chain Reaction
  • Promoter Regions, Genetic
  • Ovarian Neoplasms
  • Oncology & Carcinogenesis
  • Neoplasms, Glandular and Epithelial
  • Neoplasm Staging
  • Middle Aged
  • Humans
  • Female
 

Citation

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Bondurant, A. E., Huang, Z., Whitaker, R. S., Simel, L. R., Berchuck, A., & Murphy, S. K. (2011). Quantitative detection of RASSF1A DNA promoter methylation in tumors and serum of patients with serous epithelial ovarian cancer. Gynecol Oncol, 123(3), 581–587. https://doi.org/10.1016/j.ygyno.2011.08.029
Bondurant, Amy E., Zhiqing Huang, Regina S. Whitaker, Lauren R. Simel, Andrew Berchuck, and Susan K. Murphy. “Quantitative detection of RASSF1A DNA promoter methylation in tumors and serum of patients with serous epithelial ovarian cancer.Gynecol Oncol 123, no. 3 (December 2011): 581–87. https://doi.org/10.1016/j.ygyno.2011.08.029.
Bondurant AE, Huang Z, Whitaker RS, Simel LR, Berchuck A, Murphy SK. Quantitative detection of RASSF1A DNA promoter methylation in tumors and serum of patients with serous epithelial ovarian cancer. Gynecol Oncol. 2011 Dec;123(3):581–7.
Bondurant, Amy E., et al. “Quantitative detection of RASSF1A DNA promoter methylation in tumors and serum of patients with serous epithelial ovarian cancer.Gynecol Oncol, vol. 123, no. 3, Dec. 2011, pp. 581–87. Pubmed, doi:10.1016/j.ygyno.2011.08.029.
Bondurant AE, Huang Z, Whitaker RS, Simel LR, Berchuck A, Murphy SK. Quantitative detection of RASSF1A DNA promoter methylation in tumors and serum of patients with serous epithelial ovarian cancer. Gynecol Oncol. 2011 Dec;123(3):581–587.
Journal cover image

Published In

Gynecol Oncol

DOI

EISSN

1095-6859

Publication Date

December 2011

Volume

123

Issue

3

Start / End Page

581 / 587

Location

United States

Related Subject Headings

  • Tumor Suppressor Proteins
  • Real-Time Polymerase Chain Reaction
  • Promoter Regions, Genetic
  • Ovarian Neoplasms
  • Oncology & Carcinogenesis
  • Neoplasms, Glandular and Epithelial
  • Neoplasm Staging
  • Middle Aged
  • Humans
  • Female