Detection of breast cancer cells in ductal lavage fluid by methylation-specific PCR

Evron, Ella; Dooley, William C.; Umbricht, Christopher B.; Rosenthal, Dorothy L.; Sacchi, Nicoletta; Gabrielson, Edward; Soito, Angela B.; Hung, David T.; Ljung, Britt Marie; Davidson, Nancy E.; Sukumar, Saraswati

doi:10.1016/s0140-6736(00)04501-3

Cited by 316 publications

(200 citation statements)

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“…The majority of samples (83%) showed methylation of at least one gene and one-fifth demonstrated three or more methylated genes, indicated by an MI X0. 43.…”

Section: Resultsmentioning

confidence: 99%

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Concurrent methylation of multiple genes in childhood ALL: Correlation with phenotype and molecular subgroup

et al. 2003

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Multiple genes have been shown to be independently hypermethylated in lymphoid malignancies. We report here on the extent of concurrent methylation of E-cadherin, Dap-kinase, O 6 MGMT, p73, p16, p15 and p14 in 129 pediatric ALL cases. While most of these genes demonstrated methylation in a proportion of cases, O 6 MGMT, p16 and p14 were infrequently methylated (11, 7 and 3%, respectively). Methylation of at least one gene was found in the vast majority (83%) of cases. To determine the extent and concordance of methylation we calculated a methylation index (MI ¼ number of methylated genes/number of studied genes) for each sample. The average MI was 0.28, corresponding to 2/7 methylated genes. MI was correlated with standard prognostic factors, including immunophenotype, age, sex, WBC and presence of specific translocations (TEL-AML1, BCR-ABL, E2A-PBX1 or MLL-AF4). We determined that children X10 years old and children presenting with high WBC (X50 Â 10 9 /l) both associated with a higher MI (Po0.01 and o0.05, respectively). T-ALLs demonstrated a lower MI (median ¼ 0.17) than precursor B ALLs (median ¼ 0.28). Among the different molecular subgroups, MLL-ALLs had the highest MI (mean ¼ 0.35), while ALLs carrying the t(1;19) had the lowest MI (mean ¼ 0.07). The most common epigenetic lesion in childhood ALL was methylation of E-cadherin (72%) independent of the molecular subtype or other clinicopathological factors.

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“…The majority of samples (83%) showed methylation of at least one gene and one-fifth demonstrated three or more methylated genes, indicated by an MI X0. 43.…”

Section: Resultsmentioning

confidence: 99%

“…[43][44][45] Indeed, some promoter hypermethylation can occur early in tumor progression. Furthermore, the positive PCR signal from such a lesion will not be masked by the presence of normal cells.…”

Section: Discussionmentioning

confidence: 99%

Concurrent methylation of multiple genes in childhood ALL: Correlation with phenotype and molecular subgroup

et al. 2003

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“…Methylation at these residues was observed in 7/10 cancer cell lines, 12/20 primary breast cancers, but none of the eight normal breast epithelial cultures or eight lymphocyte DNAs (from a subset of the breast cancer patients whose tumors were analysed). Therefore, methylation at these sites appears to be a common Evron et al, 2001a). That GADD45a is hypermethylated in breast cancer does not come as a surprise.…”

Section: Discussionmentioning

confidence: 96%

“…Since hypermethylated genes may be involved in the etiology of the disease and may be early events in neoplastic progression, these genes make attractive targets for therapeutics and disease detection/monitoring (Bovenzi et al, 1999;Evron et al, 2001a;Umbricht et al, 2001;Muller et al, 2003;Oshiro et al, 2003;Parker et al, 2003). Agents that reactivate gene expression silenced by methylation have favorable therapeutic indices and are now entering clinical trials for a variety of cancers (Baylin, 2004;Dowell and Minna, 2004;Egger et al, 2004;Zelent et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Analysis of methylation-sensitive transcriptome identifies GADD45a as a frequently methylated gene in breast cancer

et al. 2005

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Treatment of the breast cancer cell line, MDAMB468 with the DNA methylation inhibitor, 5-azacytidine (5-AzaC) results in growth arrest, whereas the growth of the normal breast epithelial line DU99 (telomerase immortalized) is relatively unaffected. Comparing gene expression profiles of these two lines after 5-AzaC treatment, we identified 36 genes that had relatively low basal levels in MDAMB468 cells compared to the DU99 line and were induced in the cancer cell line but not in the normal breast epithelial line. Of these genes, 33 have associated CpG islands greater than 300 bp in length but only three have been previously described as targets for aberrant methylation in human cancer. Northern blotting for five of these genes (a-Catenin, DTR, FYN, GADD45a, and Zyxin) verified the array results. Further analysis of one of these genes, GADD45a, showed that 5-AzaC induced expression in five additional breast cancer cell lines with little or no induction in three additional lines derived from normal breast epithelial cells. The CpG island associated with GADD45a was analysed by bisulfite sequencing, sampling over 100 CpG dinucleotides. We found that four CpG's, located approximately 700 bp upstream of the transcriptional start site are methylated in the majority of breast cancer cell lines and primary tumors but not in DNA from normal breast epithelia or matched lymphocytes from cancer patients. Therefore, this simple method of dynamic transcriptional profiling yielded a series of novel methylation-sensitive genes in breast cancer including the BRCA1 and p53 responsive gene, GADD45a.

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“…These methods included FISH analysis for aneusomy or MSP for DNA methylation profile. 21,22 However, compared to FISH and MSP methods, our fluorescence image-based approach has 1 obvious advantage: the analysis is performed on the same cells on the original slide after morphological evaluation. Direct analysis of biomarker expression with morphology and analysis of association between markers on an individual cell basis is possible.…”

Section: Discussionmentioning

confidence: 99%

Biomarker analysis on breast ductal lavage cells in women with and without breast cancer

Zhang

Yang

Zhang

et al. 2006

Intl Journal of Cancer

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Recent studies show that morphology based analysis of ductal lavage specimens failed to detect many cancers in women with breast cancer. Such an observation raises doubts about the potential role of ductal lavage in an individual's risk assessment and early detection of breast cancer. We hypothesize that biomarkerbased analysis using markers of malignancy field defects including DNA 5c exceeding rate (DNA 5cER) and G-actin might provide a more reliable test for breast cancer risk. The study was performed in 2 phases, the training and validation phase. For the training phase, 36 Chinese women were recruited (13 with breast cancer, 8 with intraductal papilloma and 15 with benign breast diseases). The validation phase included 10 women with cancer and 7 women without cancer. Ductal lavage samples were processed by the ThinPrep technique and evaluated by morphology followed by biomarker analysis using laser scan cytometry (LSC) for G-actin and DNA5cER. In the training phase, biomarker analysis was performed on the 67% (24 of 36) of samples that had over 100 epithelial cells. The sensitivity of DNA5cER was 90% with a specificity of 100%, and G-actin was 100% and 93%, respectively. By contrast, the sensitivity and specificity obtained by cytology alone were 67% and 93%, respectively. Similar results were obtained from the small validation study. Quantitative analysis of biomarkers for G-actin and DNA5cER is feasible and useful in distinguishing benign from malignant breast disease on archived ductal lavage slides. Further studies are warranted to determine the value of these biomarkers in prospective trials. ' 2006 Wiley-Liss, Inc.

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Detection of breast cancer cells in ductal lavage fluid by methylation-specific PCR

Cited by 316 publications

References 4 publications

Concurrent methylation of multiple genes in childhood ALL: Correlation with phenotype and molecular subgroup

Concurrent methylation of multiple genes in childhood ALL: Correlation with phenotype and molecular subgroup

Analysis of methylation-sensitive transcriptome identifies GADD45a as a frequently methylated gene in breast cancer

Biomarker analysis on breast ductal lavage cells in women with and without breast cancer

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