Loss of Heterozygosity Analysis in Ductal Lavage Samples from <i>BRCA1</i> and <i>BRCA2</i> Carriers: A Cautionary Tale

Antill, Yoland

doi:10.1158/1055-9965.epi-05-0986

Cited by 5 publications

(6 citation statements)

References 15 publications

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“…30,31). Although the rate of epithelial CN/ LOH events in this study is significantly lower than previously reported, artifactual LOH events may have been introduced when conducting microsatellite analyses on low quantities of DNA in these previous studies (32). Our findings are consistent with the findings of Cheng and colleagues (33) who showed in a cohort of 29 ovarian serous cystadenomas that only 14% of these tumors had detectable monoclonal expansion in the epithelium.…”

Section: Discussionsupporting

confidence: 90%

Copy Number Aberrations in Benign Serous Ovarian Tumors: A Case for Reclassification?

Hunter

Anglesio

Sharma

et al. 2011

Clinical Cancer Research

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Purpose: Serous ovarian carcinomas are the predominant epithelial ovarian cancer subtype and it has been widely believed that some or all of these may arise from precursors derived from the ovarian surface epithelium or fimbriae, although direct molecular evidence for this is limited. This study aimed to conduct copy number (CN) analysis using a series of benign and borderline serous ovarian tumors to identify underlying genomic changes that may be indicative of early events in tumorigenesis.Experimental Design: High resolution CN analysis was conducted on DNA from the epithelial and fibroblast components of a cohort of benign (N ¼ 39) and borderline (N ¼ 24) serous tumors using the Affymetrix OncoScan assay and SNP6.0 arrays.Results: CN aberrations were detected in the epithelium of only 2.9% (1 of 35) of serous cystadenomas and cystadenofibromas. In contrast, CN aberrations were detected in the epithelium of 67% (16 of 24) of the serous borderline tumors (SBT). Unexpectedly, CN aberrations were detected in the fibroblasts of 33%

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Section: Discussionsupporting

confidence: 90%

Copy Number Aberrations in Benign Serous Ovarian Tumors: A Case for Reclassification?

Hunter

Anglesio

Sharma

et al. 2011

Clinical Cancer Research

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“…If loss of the wildtype allele at the relevant BRCA locus in germ-line BRCA mutation carriers represents an early event in breast tumorigenesis, analysis of free DNA from ductal lavage fluid may offer a useful alternative surrogate marker of risk particularly where samples are insufficient for cytologic diagnosis. However, the degree of reproducibility of these assays found in this study and that of Antill et al (13) raises questions about the potential of this technique as a robust risk assessment tool in the evaluation of women with a genetic predisposition to breast cancer. Further studies are required to optimize the reproducibility of the methods and to evaluate the specificity and predictive value of LOH in ductal lavage fluid for subsequent breast cancer development.…”

Section: Resultsmentioning

confidence: 57%

Loss of Heterozygosity at theBRCA1andBRCA2Loci Detected in Ductal Lavage Fluid fromBRCAGene Mutation Carriers and Controls

Locke

Kóte-Jarai

Bancroft

et al. 2006

Cancer Epidemiology, Biomarkers &Amp; Prevention

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Female BRCA gene mutation carriers are at increased risk for developing breast cancer. Ductal lavage is a novel method for sampling breast ductal fluid, providing epithelial cells for cytologic assessment and a source of free DNA for molecular analyses. Loss of heterozygosity (LOH) at the BRCA loci in ductal lavage fluid is a potential biomarker of breast cancer risk. The LOH rate was measured at the BRCA1/2 loci and compared with that at a control locus (APC) using free DNA from the ductal lavage fluid of BRCA carriers and predictive test negative controls. We evaluated the reproducibility of these analyses. Free DNA sufficient for PCR amplification was obtained from 33 ductal lavage samples of 17 healthy women of known BRCA status (14 BRCA carriers and 3 controls). LOH rates of 36.4% to 56.3% at the BRCA1 locus and 45% to 61.5% at the BRCA2 locus were found among BRCA carriers. The LOH rate at the APC locus was lower (18.5%). The interaliquot reproducibility for the D17S855 marker of the BRCA1 locus was 66.7%. Intraaliquot reproducibility was 90%. Although we successfully isolated sufficient free DNA from ductal lavage fluid for PCR amplification, the degree of reproducibility of these LOH studies raises questions about the robustness of this technique as a risk assessment tool in the evaluation of high-risk women. Further studies are required to evaluate the specificity and predictive value of LOH in ductal lavage fluid for breast cancer development.

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“…DL, nipple aspiration, and venesection were completed every 6 mo from the time of study for a maximum of 3 y with annual clinical follow-up planned for 10 y, again as previously described (21,22).…”

Section: Specimen Collectionmentioning

confidence: 99%

“…Between 10 and 12 mL of DL, washings were recovered from each duct with processing of the sample within 2 h by tissue bank staff as previously described (21,22). Briefly, DL fluid was spun before removal of the supernatant down to 2 mL.…”

Section: Sample Processingmentioning

confidence: 99%

“…DNA was extracted from DL cellular pellets using the DNeasy Tissue kit (Qiagen), and lymphocytic DNA was processed using the Mini Blood kit (Qiagen). The DNA yield was estimated for a subset of samples using quantitative PCR, with DNA quantity estimated by plotting against a standard curve of known dilutions of DNA derived from normal lymphocytes as described previously (22,23).…”

Section: Sample Processingmentioning

confidence: 99%

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Gene Methylation in Breast Ductal Fluid from BRCA1 and BRCA2 Mutation Carriers

Antill

Mitchell²,

Johnson³

et al. 2010

Cancer Epidemiology, Biomarkers &Amp; Prevention

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Purpose: Genomic alterations (including gene hypermethylation) are likely to precede the phenotypic changes associated with breast tumorigenesis. From a prospective collection of ductal lavage (DL) samples from women with a known mutation in BRCA1 or BRCA2, we have assessed promoter methylation with a comparison of results with several variables, including breast cancer (BC) outcome.Experimental Design: Hypermethylation of p16, RASSF1A, twist, and RARβ was assessed using a qualitative, real-time, nested PCR assay. Associations between methylation status and variables were tested using Fisher's exact test or logistic regression. Analyses were done at three levels: a single breast, a single duct (both over time), and each DL sample in isolation.Results: A total of 168 samples from 93 ducts in 54 breasts have been analyzed in 34 women (16 BRCA1 and 18 BRCA2 mutation carriers). A median of 2 DL was done (range, 1-5), with 7 women developing BC on study, 1 bilateral. Methylation of p16 was associated with a known BRCA1 mutation (P = 0.001, P < 0.001, and P < 0.001 for breast, duct, and sample levels, respectively) and women with a history of contralateral BC (P = 0.001 and P < 0.001 for duct and sample levels, respectively). An association was seen for women who developed BC on study and RASSF1A methylation (P = 0.001 for sample level).Conclusions: Genetic methylation patterns could potentially be used to predict future BC risk. In addition, p16 methylation may be a predictor of BRCA1 mutation status. Further research is required to corroborate these findings.

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Loss of Heterozygosity Analysis in Ductal Lavage Samples from BRCA1 and BRCA2 Carriers: A Cautionary Tale

Cited by 5 publications

References 15 publications

Copy Number Aberrations in Benign Serous Ovarian Tumors: A Case for Reclassification?

Copy Number Aberrations in Benign Serous Ovarian Tumors: A Case for Reclassification?

Loss of Heterozygosity at theBRCA1andBRCA2Loci Detected in Ductal Lavage Fluid fromBRCAGene Mutation Carriers and Controls

Gene Methylation in Breast Ductal Fluid from BRCA1 and BRCA2 Mutation Carriers

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