Annals of Surgical Oncology Sign the Guestbook
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Originally published as Ann Surg Oncol Early Release 10.1245/ASO.2004.03.024 on August 16, 2004

Annals of Surgical Oncology 11:861-868 (2004)
© 2004 Society of Surgical Oncology
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ellsworth, D. L.
Right arrow Articles by Shriver, C. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ellsworth, D. L.
Right arrow Articles by Shriver, C. D.

ORIGINAL ARTICLES

Outer Breast Quadrants Demonstrate Increased Levels of Genomic Instability

Darrell L. Ellsworth, PhD, Rachel E. Ellsworth, PhD, Brad Love, PhD, Brenda Deyarmin, Susan M. Lubert, Vimal Mittal, MD, Jeffrey A. Hooke, MD and Craig D. Shriver, MD

From the Clinical Breast Care Project, Windber Research Institute (DLE, REE, BD, SML, VM), Windber, Pennsylvania; Invitrogen Bioinformatics (BL), Frederick, Maryland; and General Surgery Service, Walter Reed Army Medical Center (JAH, CDS), Washington, DC.

Correspondence: Address correspondence and reprint requests to: Darrell L. Ellsworth, PhD, Windber Research Institute, 600 Somerset Avenue, Windber, PA 15963; Fax: 814-467-6334; E-mail: d.ellsworth{at}wriwindber.org


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 CONCLUSIONS
 REFERENCES
 
Background: Theory holds that the upper outer quadrant of the breast develops more malignancies because of increased tissue volume. This study evaluated genomic patterns of loss of heterozygosity (LOH) and allelic imbalance (AI) in non-neoplastic tissues from quadrants of diseased breasts following mastectomy to characterize relationships between genomic instability and the propensity for tumor development.

Methods: Tissues from breast quadrants were collected from 21 patients with various stages of breast carcinoma. DNA was isolated from non-neoplastic tissues using standard methods and 26 chromosomal regions commonly deleted in breast cancer were examined to assess genomic instability.

Results: Genomic instability was observed in breast quadrants from patients with ductal carcinomas in situ and advanced carcinomas. Levels of instability by quadrant were not predictive of primary tumor location (P = .363), but outer quadrants demonstrated significantly higher levels of genomic instability than did inner quadrants (P = .017). Marker D8S511 on chromosome 8p22–21.3, one of the most frequently altered chromosomal regions in breast cancer, showed a significantly higher level of instability (P = .039) in outer compared with inner quadrants.

Conclusions: Non-neoplastic breast tissues often harbor genetic changes that can be important to understanding the local breast environment within which cancer develops. Greater genomic instability in outer quadrants can partially explain the propensity for breast cancers to develop there, rather than simple volume-related concepts. Patterns of field cancerization in the breast appear to be complex and are not a simple function of distance from a developing tumor.

Key Words: Loss of heterozygosity • Allelic imbalance • Breast cancer • Field cancerization • Quadrants • Metastasis


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 CONCLUSIONS
 REFERENCES
 
Breast cancer is the most common cancer in women from western countries, with overall mortality surpassed only by mortality attributable to lung cancer. Breast cancer incidence has risen in the United States approximately 1.2% per year since 1930. Currently, one of eight American women can be expected to develop breast cancer during her lifetime.1 A number of prognostic factors, including hormonal status, tumor size, and presence of lymph node metastasis, have been identified to guide treatment and predict clinical outcome. Efforts to utilize tumor location as a prognostic factor have shown that breast tumors occur most frequently in the upper outer quadrant (UOQ) (48% to 54%), and approximately 66% of tumors occur in the outer quadrants compared with the inner quadrants (22%) or central region (12%).2,3 The propensity for tumor development is believed to be directly proportional to the amount of tissue in each breast quadrant, but this hypothesis of tumor development based solely on tissue volume may not accurately account for the overabundance of tumors in the outer quadrants. In hereditary colorectal cancer, for example, tumors in the smaller distal colon are associated with mutations in the adenomatous polyposis coli (APC) gene, whereas those in the larger proximal colon are associated with mutations in mismatch-repair genes.4 Similarly, genomic alterations within the breast can influence the site of tumor formation.

Genomic instability is essential for the development of sequential genetic changes that drive tumorigenic processes (e.g., neoplastic cell growth, tumor heterogeneity, tissue invasion, and metastasis),5 but little is known about either the timing of critical mutations in early carcinogenesis or the significance of genetic alterations in preneoplastic tissues. Genetic alterations known as loss of heterozygosity (LOH) and allelic imbalance (AI) have been observed in benign breast disease, including fibrocystic change (FCC),6 epithelial hyperplasia,7 and fibroadenomas,8 as well as in situ carcinomas.9 In addition, genomic instability has been documented in breast tissues that appear normal on histologic examination.10–14 Despite recent findings suggesting functional significance of somatic mutations in fibroblastic stroma,15,16 the extent of genomic instability in non-neoplastic tissues throughout the breast has not been well characterized and relationships between the location of genetic alterations in morphologically normal breast tissue and the propensity for tumor development remain unknown.

In this study, we examined genomic instability (loss of heterozygosity or allelic imbalance) in breast quadrants from 21 patients with breast carcinomas. We surveyed 52 microsatellite markers defining 26 chromosomal regions throughout the genome to (1) assess genomic instability in non-neoplastic tissues of diseased breasts following mastectomy; (2) compare levels of genomic instability in breast quadrants, both proximal to and distant from the primary tumor, with levels of genomic instability previously observed in the primary tumors and neighboring tissue from the same breast; (3) determine relationships between genomic instability in breast quadrants and patterns of tumor development; and (4) identify chromosomal regions showing frequent instability that may contain genes involved in early tumorigenic processes. We hypothesized that genomic instability in breast quadrants can be associated with the propensity for tumor development and may identify important molecular alterations associated with breast disease.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 CONCLUSIONS
 REFERENCES
 
Sample Collection and Processing
Histology
The Institutional Review Board of the Windber Medical Center approved this research. Breast tissues were surgically removed from patients during mastectomy and stored in the pathology archives of the Windber Medical Center. For each breast quadrant, representative tissue sections were embedded in paraffin, sliced at 4 µm, mounted on glass microscope slides, and then stained with hematoxylin and eosin. A board-certified pathologist viewed the slides from each patient to confirm histology and guide microdissection. Sections composed mainly of histologically normal ducts and stromal fibroblasts, and free from atypical ductal hyperplasia (ADH) or residual tumor [ductal carcinoma in situ (DCIS) or invasive carcinoma], were cut at ~10 µm and three to six slices (25 mg) were used for DNA isolation. For blocks containing ADH or residual tumor, 16 serial sections (5 µm) were cut, and an ASLMD Laser Microdissection system (Leica Microsystems, Wetzlar, Germany) was used to extract normal-appearing ducts and stromal cells, avoiding areas of breast disease.17

Quadrants were collected from 21 patients at various stages of breast disease, including premalignant DCIS (n = 4) and invasive ductal carcinomas, stage I (n = 6), stage II (n = 7), and stage III (n = 4). All tumors were staged using the tumor, node, metastasis staging system approved by the American Joint Commission on Cancer, 6th ed.18 The in situ carcinomas were classified by cytonuclear grade and presence or absence of necrosis.19 For the invasive lesions, the degree of tumor differentiation was assessed by Scarff-Bloom-Richardson (SBR) grading, which evaluated tubule formation, nuclear pleomorphism, and mitotic index.20 All breast quadrant samples were characterized histologically to determine presence of FCC, which included primarily sclerosing adenosis and apocrine metaplasia.

DNA Isolation
For sections free from residual breast disease, DNA was isolated using the QIAamp DNA Mini Kit (QIAGEN, Valencia, CA). DNA was extracted from laser microdissected material by overnight digestion in proteinase K (0.4 mg/mL) at 37°C and purified by centrifugation at 10,000x g through Microcon YM-50 centrifugal filters (Millipore, Bedford, MA). For each sample, normal tissue free from any histologic abnormalities and distant from sites of tissue collection (e.g., negative axillary lymph node or normal nipple or skin tissue) was used as a source of referent DNA.

Genotyping
We used a custom panel of 52 microsatellite markers (Invitrogen, Carlsbad, CA) representing 26 chromosomal regions commonly deleted in breast cancer to assess patterns of genomic instability throughout the genome. Polymerase chain reaction (PCR) was conducted with puReTaq Ready-To-Go PCR beads (Amersham Biosciences, Buckinghamshire, England) or AmpliTaq Gold PCR Master Mix (Applied Biosystems, Foster City, CA) using amplification conditions previously described.21 Following amplification, samples were purified using Sephadex G-50 resin (Amersham Biosciences) and then genotyped on a MegaBACE 1000 capillary electrophoresis DNA analysis system (Amersham Biosciences) using Genetic Profiler software (version 1.5).

Statistical Analysis
The degree of chromosomal loss at each microsatellite marker was estimated by calculating normalized ratios of allele peak heights using the formula (T1/T2)/(N1/N2) where T1 and N1 are the smaller peak heights in the quadrant and referent samples, respectively, and T2 and N2 are the larger peak heights in the quadrant and referent samples, respectively.22,23 Any quadrant with a normalized ratio of ≤0.35 was considered to show genomic instability, which indicates that a substantial proportion of cells in that quadrant sample contained the same chromosomal aberration compared with normal somatic cells. A normalized ratio of ≤0.35 can result from loss (of heterozygosity) or an increase in copy number (allelic amplification) of a chromosomal region.

We compared the level of genomic instability (1) among all breast quadrants, (2) between inner versus outer and upper versus lower quadrants, and (3) in quadrants relative to that in the primary tumor and in tissue immediately adjacent to the tumor, using Fisher’s exact test (one-sided) under a hypergeometric distribution. A {chi}2 test for randomness was used to determine if the extent of genomic instability in each breast quadrant was predictive of tumor location or, in other words, to determine if genomic instability was observed more frequently in the quadrant where the primary tumor was located. Fisher’s exact test was also used to assess patterns of genomic instability across all markers, as well as relationships between genomic instability and (1) the distribution of FCC throughout the breast and (2) presence of regional metastasis to the axillary lymph nodes.

Quality Control
Histologic examination of all tissue sections and laser microdissection of quadrant samples containing residual tumor were conducted in close collaboration with the pathologist to ensure consistency in the clinical diagnoses and to accurately identify diseased cells. The same areas of tissue were microdissected across multiple sections to minimize possible regional differences in patterns of genomic instability.24 A minimal acceptable signal intensity in the electropherograms (1000 relative fluorescence units) was implemented to increase the accuracy of the allele calls. Chromosomal regions showing evidence of instability were assayed a second time in an independently microdissected sample when sufficient tissue was available. For all replicate assays, the average of the normalized ratios for the independent assays was used in the statistical analyses.

Representative results were rigorously examined for stochastic artifactual variation by testing for correlations between DNA concentration, percentage of missing genotypes, presence of residual tumor in the section (removed by laser microdissection), and percentage of informative markers showing genomic instability. Contingency tables and Fisher’s exact test for independence with a continuity correction were used to determine significance of these comparisons. In addition, a binomial probability (two-tailed) test was used to examine ratios of large versus small alleles altered at sites of genomic instability. Finally, to establish the background level of genomic instability in normal breast tissue from disease-free patients, paraffin-embedded samples were obtained from three reduction mammoplasty cases and processed as described above. Peak heights from dense fibrous mesenchymal tissue were compared with those in skin (epidermal through dermal layer) or peripheral blood to assess genomic instability.


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 CONCLUSIONS
 REFERENCES
 
In this study, the primary tumor was located most frequently in the lower outer quadrant (30.9%) and least frequently in the upper inner quadrant (11.9%) (Table 1). Levels of genomic instability in tissues located within the same quadrant as the primary tumor, but not immediately adjacent to the tumor, did not differ significantly (P = .363) from levels of instability in the other quadrants of a given breast, which indicates that levels of genomic instability were not predictive of primary tumor location.


View this table:
[in this window]
[in a new window]
  		TABLE 1. Primary tumor location stratified by breast quadrant
 
The primary tumor (P < .0001) and adjacent stroma (P < .05) showed a significantly higher level of genomic instability than any of the quadrants (Table 2). When quadrant samples were combined to compare upper versus lower and inner versus outer quadrants, genomic instability did not differ (P = .456) between upper versus lower quadrants, but was significantly higher (P = .017) in the outer quadrants relative to the inner quadrants (Fig. 1). Across all 26 chromosomal regions, 17 regions showed relatively more genomic instability in outer quadrants relative to inner quadrants, whereas only three regions demonstrated greater instability in the inner quadrants (Fig. 2). Marker D8S511 on chromosome 8p showed a significant difference in instability (P = .039) between inner and outer quadrants.


View this table:
[in this window]
[in a new window]
  		TABLE 2. Average frequency (%) of genomic instability per patient in primary tumor, adjacent stroma, and breast quadrant samples
 


View larger version (12K):
[in this window]
[in a new window]
  		FIG. 1. Average frequency of genomic instability per patient in breast quadrant samples.

 


View larger version (25K):
[in this window]
[in a new window]
  		FIG. 2. Levels of genomic instability across 26 chromosomal regions commonly deleted in breast cancer for outer versus inner quadrants. Marker D8S511 at chromosome 8p22–21.3 (indicated by an asterisk) showed significantly greater genomic instability in outer versus inner quadrants.

 
Distribution of FCC throughout the breast did not differ significantly between upper versus lower (P = 1.000) or inner versus outer (P = .239) quadrants. In upper and outer breast quadrants, the degree of genomic instability was inversely related to presence of FCC: quadrants with FCC showed significantly lower levels of genomic instability (P< .001) (Table 3). When patients were stratified by axillary lymph node status, genomic instability was significantly higher in lower (P < .001) and outer (P < .001) quadrants in patients with regional metastasis (Table 4).


View this table:
[in this window]
[in a new window]
  		TABLE 3. Relationship between genomic instabilitya and fibrocystic change in breast quadrants
 

View this table:
[in this window]
[in a new window]
  		TABLE 4. Relationship between genomic instabilitya in breast quadrants and regional metastasis to the axillary lymph nodes
 
Quality control analyses suggest that no systematic artifactual variation was present in the data. No statistical correlations were detected between presence of residual tumor and percentage of markers showing genomic instability (P = .645). Likewise, the DNA concentration of each sample was not associated with missing genotypes (P = .927) or the percentage of markers showing evidence of instability (P = .326). The minimal amount of DNA per amplification that produced repeatable results was 2 ng. A relationship was seen between presence of residual tumor, hence the need for laser microdissection, and percentage of missing genotypes (P < .01), with laser microdissected samples producing a greater number of missing genotypes. For markers showing evidence of genomic instability, large alleles were not altered significantly more often than small alleles (P = .226). The average frequency of genomic instability per patient in the reduction mammoplasty samples was <0.8%, demonstrating that our methods detected a very low level of genomic instability in disease-free breast tissue.


   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
CONCLUSIONS
 REFERENCES
 
The concept of field cancerization was originally proposed by Slaughter et al.25 in 1953 to explain multiple independent lesions and local recurrence of oral cancer. Under the field cancerization hypothesis, the propensity for tumor development is greatly increased in specific areas of tissue because of genetic alterations resulting from long-term exposure to carcinogens. Recent molecular findings for many types of cancer, including head and neck squamous cell carcinoma,26,27 lung,28 esophagus,29 cervix,30 colon,31 and breast,10 suggest that fields of genetically altered cells can develop from single stem cells that have acquired specific genetic changes and, thus, have escaped normal growth control. The proliferating preneoplastic fields, which show normal histology or features characteristic of dysplasia, gradually displace native "genetically normal" tissues locally. With the accumulation of additional genetic alterations, normal appearing cells may give rise to in situ carcinomas and eventually invasive cancer.32

The frequent occurrence of carcinomas in outer breast quadrants was traditionally believed to be a simple function of tissue volume. In this study, however, we observed greater genomic instability in outer breast quadrants compared with the inner quadrants, which may partially explain the increased tendency for tumors to develop in the outer breast hemisphere. Genomic instability was not related to benign tissue changes (e.g, sclerosing adenosis or apocrine metaplasia). Although genetic alterations (LOH) have been observed in components of FCC,6 FCC is often included within the spectrum of normal physiologic variation.33 We believe the increased level of genomic instability in the outer quadrants reflects innate instability of the local tissues, rather than instability attributable to lymphogenic spread of tumor cells from the primary carcinoma, because (1) we would expect the relative number of migrating tumor cells to be very low in each tissue section and (2) the patterns of LOH/AI were unique to each quadrant and differed from those observed in the primary tumor.

We hypothesize that increased genomic instability in outer breast quadrants can be attributable to field cancerization effects. The reasons behind this breast field cancerization effect are unclear at this point; perhaps it results from lymphatic drainage in the breast and exposure to carcinogens. Lymphatic mapping of breast drainage patterns has shown that drainage toward the axillary lymph nodes through the outer quadrants occurs more frequently than drainage through the internal mammary chain region.34 As the lymphatic system transports waste products of metabolism, a more extensive drainage system in the outer quadrants may increase exposure of tissues in this region to carcinogens and contribute to greater genomic instability.

Frequent loss of heterozygosity at specific chromosomal regions in certain cancers implies the presence of tumor suppressor genes. The commonly deleted region on chromosome 8p22–21.3 frequently shows LOH in a number of human epithelial cancers,35 and may be relevant to early breast cancer development because this region demonstrates allelic loss in early stage prostate cancer36 and appears to be one of the most frequently altered chromosomal regions in breast cancer.37 Genomic instability at chromosome 8p22–21.3 is common in preinvasive in situ carcinomas,38 but this region has not been well studied in early stages of breast disease. Several genes localized to the 8p region with a putative role in cancer development are shown in Table 5.39–44 Of note, the DLC-1 gene shows reduced levels of expression in 70% of breast carcinoma cell lines and has a significant inhibitory effect on tumorigenicity in mice.39 Further study is needed to determine if genetic alterations in one or more genes located at 8p22–21.3 in preneoplastic tissues contribute to carcinogenesis.


View this table:
[in this window]
[in a new window]
  		TABLE 5. Genes with a putative role in cancer development localized close to the region on chromosome 8p22-21.3a
 
Multiple studies have found associations between allelic loss at specific chromosomal regions in primary breast carcinomas and clinical outcomes (e.g., regional metastasis),45–47 but little is known about overall patterns of genomic instability throughout the breast and axillary lymph node involvement. It is widely accepted that metastatic axillary lymph node involvement is associated with tumors located in outer quadrants.3,48 Our observation that levels of genomic instability in outer breast quadrants are significantly higher in patients who are node-positive compared with those who are node-negative reinforces the notion that genomic instability in outer quadrants is associated with more frequent carcinogenesis in the outer quadrants and ensuing axillary lymph node involvement.

Local recurrence of breast cancer is highly prevalent (~73%) in the same quadrant as the previous excision49 and traditionally has been attributed to incomplete resection of the primary carcinoma. An emerging alternative hypothesis that may partially explain local recurrence within the same quadrant is the presence of a genetically altered, preneoplastic field left behind after surgery. Evidence for the existence of genetically altered preneoplastic fields in breast cancer is now being recognized as a possible explanation of recurrence for both ductal carcinoma in situ50 and invasive carcinomas.51 We observed significantly lower levels of genomic instability at the biopsy site, which indicates that tissue in the immediate vicinity of the primary tumor may not be genetically altered, but residual genomic instability can occur in non-neoplastic tissues distant from the site of the primary tumor. Current definitions of non-neoplastic tissues, thus, need refinement based on both morphologic and genetic information.

Genetically altered preneoplastic fields are difficult to recognize clinically, and histologic examination has not proved to be a reliable method for identifying lesions at risk for progression. The ability to link specific patterns of genetic alteration to disease progression has great potential for improving clinical detection of breast cancer and guiding treatment. Preliminary studies of oral squamous cell carcinomas indicate that preneoplastic lesions progressing to malignancy show significantly greater genomic instability (AI) than similar lesions that do not progress52 and allelic loss (LOH) at specific chromosomal regions may be useful in predicting cases susceptible to progression.53 Although breast quadrant sections have been shown to provide information on multifocality, vascular invasion, and margin involvement,54 an assessment of genomic instability in quadrant sections may provide valuable predictive information regarding metastasis and local recurrence after breast conserving therapy.


   CONCLUSIONS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 CONCLUSIONS
REFERENCES
 
Breast tissues that appear non-neoplastic pathologically often harbor genetic changes that can be important to understanding the local breast environment within which cancer develops. In this study, we observed greater genomic instability in outer quadrants of the breast, which can partially explain the propensity for breast cancers to develop there, rather than simple volume-related breast tissue concepts. Our data further suggest that patterns of field cancerization in the breast are complex—not a simple function of distance from a developing tumor. These findings have implications for predicting regional metastasis and local recurrence. Additional research is needed to identify molecular signatures of breast disease that identify genetically altered preneoplastic tissues and are predictive of breast cancer development and recurrence.


   ACKNOWLEDGMENTS
 
The Clinical Breast Care Project (CBCP) is supported by the United States Department of Defense and the Henry M. Jackson Foundation for the Advancement of Military Medicine (MDA 905–00–1–0022). The joint efforts of many CBCP investigators and staff members are gratefully acknowledged. John F. Yerger, Harold Ashcraft, and the staff of the Windber Medical Center Pathology Department provided access to archival breast samples and contributed invaluable guidance. The opinion and assertions contained herein are the private views of the authors and are not to be construed as official or as representing the views of the Department of the Army or the Department of Defense.


   FOOTNOTES
 
Greater innate genomic instability can partially explain the preferential development of breast cancers in outer breast quadrants. Patterns of field cancerization in the breast appear to be complex and are not a simple function of distance from a developing tumor.

Received for publication March 18, 2004. Accepted for publication May 12, 2004.


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 CONCLUSIONS
 REFERENCES
 

  1. Jemal A, Murray T, Samuels A, Ghafoor A, Ward E, Thun MJ. Cancer statistics, 2003. CA Cancer J Clin 2003; 53: 5–26.[Abstract/Free Full Text]
  2. Bevilacqua JLB, Cody HS III, MacDonald KA, Tan LK, Borgen PI, Van Zee KJ. A model for predicting axillary node metastases based on 2000 sentinel node procedures and tumour position. Eur J Surg Oncol 2002; 28: 490–500.[CrossRef][Medline]
  3. Janni W, Rack B, Sommer H, et al. Intra-mammary tumor location does not influence prognosis but influences the prevalence of axillary lymph-node metastases. J Cancer Res Clin Oncol 2003; 129: 503–10.[CrossRef][Medline]
  4. Lynch HT, de la Chapelle A. Hereditary colorectal cancer. N Engl J Med 2003; 348: 919–32.[Free Full Text]
  5. Lengauer C, Kinzler KW, Vogelstein B. Genetic instabilities in human cancers. Nature 1998; 396: 643–9.[CrossRef][Medline]
  6. Washington C, Dalbègue F, Abreo F, Taubenberger JK, Lichy JH. Loss of heterozygosity in fibrocystic change of the breast: genetic relationship between benign proliferative lesions and associated carcinomas. Am J Pathol 2000; 157: 323–9.[Abstract/Free Full Text]
  7. Kaneko M, Arihiro K, Takeshima Y, Fujii S, Inai K. Loss of heterozygosity and microsatellite instability in epithelial hyperplasia of the breast. J Exp Ther Oncol 2002; 2: 9–18.[CrossRef][Medline]
  8. Tibiletti MG, Sessa F, Bernasconi B, et al. A large 6q deletion is a common cytogenetic alteration in fibroadenomas, pre-malignant lesions, and carcinomas of the breast. Clin Cancer Res 2000; 6: 1422–31.[Abstract/Free Full Text]
  9. O’Connell P, Pekkel V, Fuqua SA, Osborne CK, Clark GM, Allred DC. Analysis of loss of heterozygosity in 399 premalignant breast lesions at 15 genetic loci. J Natl Cancer Inst 1998; 90: 697–703.[Abstract/Free Full Text]
  10. Deng G, Lu Y, Zlotnikov G, Thor AD, Smith HS. Loss of heterozygosity in normal tissue adjacent to breast carcinomas. Science 1996; 274: 2057–9.[Abstract/Free Full Text]
  11. Larson PS, de las Morenas A, Cupples LA, Huang K, Rosenberg CL. Genetically abnormal clones in histologically normal breast tissue. Am J Pathol 1998; 152: 1591–8.[Abstract]
  12. Moinfar F, Man YG, Bratthauer GL, Ratschek M, Tavassoli FA. Genetic abnormalities in mammary ductal intraepithelial neoplasia-flat type ("clinging ductal carcinoma in situ"): a simulator of normal mammary epithelium. Cancer 2000; 88: 2072–81.[CrossRef][Medline]
  13. Försti A, Louhelainen J, Söderberg M, Wijkström H, Hemminki K. Loss of heterozygosity in tumour-adjacent normal tissue of breast and bladder cancer. Eur J Cancer 2001; 37: 1372–80.
  14. Kurose K, Hoshaw-Woodard S, Adeyinka A, Lemeshow S, Watson PH, Eng C. Genetic model of multi-step breast carcinogenesis involving the epithelium and stroma: clues to tumour-microenvironment interactions. Hum Mol Genet 2001; 10: 1907–13.[Abstract/Free Full Text]
  15. Wernert N, Locherbach C, Wellmann A, Behrens P, Hugel A. Presence of genetic alterations in microdissected stroma of human colon and breast cancers. Anticancer Res 2001; 21: 2259–64.[Medline]
  16. Kurose K, Gilley K, Matsumoto S, Watson PH, Zhou X-P, Eng C. Frequent somatic mutations in PTEN and TP53 are mutually exclusive in the stroma of breast carcinomas. Nat Genet 2002; 32: 355–7.[CrossRef][Medline]
  17. Ellsworth DL, Shriver CD, Ellsworth RE, Deyarmin B, Somiari RI. Laser capture microdissection of paraffin-embedded tissues. Biotechniques 2003; 34: 42–6.[Medline]
  18. American Joint Committee on Cancer. AJCC Cancer Staging Manual. 6th ed. New York: Springer, 2002.
  19. Millis RR, Hanby AM, Oberman HA. The breast. In: Sternberg SS, ed. Diagnostic Surgical Pathology. 3rd ed. Philadelphia: Lippincott, Williams & Wilkins, 1999; 319–85.
  20. Elston CW, Ellis IO. Pathological prognostic factors in breast cancer. I. The value of histological grade in breast cancer: experience from a large study with long-term follow-up. Histopathology 1991; 19: 403–10.[Medline]
  21. Ellsworth RE, Ellsworth DL, Lubert SM, Hooke J, Somiari RI, Shriver CD. High-throughput loss of heterozygosity mapping in 26 commonly deleted regions in breast cancer. Cancer Epidemiol Biomarkers Prev 2003; 12: 915–9.[Abstract/Free Full Text]
  22. Wang Y, Hung S-C, Linn JF, et al. Microsatellite-based cancer detection using capillary array electrophoresis and energy-transfer fluorescent primers. Electrophoresis 1997; 18: 1742–9.[CrossRef][Medline]
  23. Medintz IL, Lee C-CR, Wong WW, Pirkola K, Sidransky D, Mathies RA. Loss of heterozygosity assay for molecular detection of cancer using energy-transfer primers and capillary array electrophoresis. Genome Res 2000; 10: 1211–8.[Abstract/Free Full Text]
  24. Wild P, Knuechel R, Dietmaier W, Hofstaedter F, Hartmann A. Laser microdissection and microsatellite analyses of breast cancer reveal a high degree of tumor heterogeneity. Pathobiology 2000; 68: 180–90.[CrossRef][Medline]
  25. Slaughter DP, Southwick HW, Smejkal W. Field cancerization in oral stratified squamous epithelium; clinical implications of multicentric origin. Cancer 1953; 6: 963–8.[CrossRef][Medline]
  26. Tabor MP, Brakenhoff RH, van Houten VMM, et al. Persistence of genetically altered fields in head and neck cancer patients: biological and clinical implications. Clin Cancer Res 2001; 7: 1523–32.[Abstract/Free Full Text]
  27. Tabor MP, Brakenhoff RH, Ruijter-Schippers HJ, et al. Multiple head and neck tumors frequently originate from a single preneoplastic lesion. Am J Pathol 2002; 161: 1051–60.[Abstract/Free Full Text]
  28. Franklin WA, Gazdar AF, Haney J, et al. Widely dispersed p53 mutation in respiratory epithelium. A novel mechanism for field carcinogenesis. J Clin Invest 1997; 100: 2133–7.[Medline]
  29. Prevo LJ, Sanchez CA, Galipeau PC, Reid BJ. p53-mutant clones and field effects in Barrett’s esophagus. Cancer Res 1999; 59: 4784–7.[Abstract/Free Full Text]
  30. Chu T-Y, Shen C-Y, Lee H-S, Liu H-S. Monoclonality and surface lesion-specific microsatellite alterations in premalignant and malignant neoplasia of uterine cervix: a local field effect of genomic instability and clonal evolution. Genes Chromosomes Cancer 1999; 24: 127–34.[CrossRef][Medline]
  31. Jothy S, Slesak B, Harlozinska A, Lapinska J, Adamiak J, Rabczynski J. Field effect of human colon carcinoma on normal mucosa: relevance of carcinoembryonic antigen expression. Tumour Biol 1996; 17: 58–64.[Medline]
  32. Braakhuis BJM, Tabor MP, Kummer JA, Leemans CR, Brakenhoff RH. A genetic explanation of Slaughter’s concept of field cancerization: evidence and clinical implications. Cancer Res 2003; 63: 1727–30.[Abstract/Free Full Text]
  33. Kamel OW, Kempson RL, Hendrickson MR. In situ proliferative epithelial lesions of the breast. Pathology 1992; 1: 65–102.[Medline]
  34. Estourgie SH, Nieweg OE, Valdés Olmos RA, Rutgers EJT, Kroon BBR. Lymphatic drainage patterns from the breast. Ann Surg 2004; 239: 232–7.[CrossRef][Medline]
  35. Arbieva ZH, Banerjee K, Kim SY, et al. High-resolution physical map and transcript identification of a prostate cancer deletion interval on 8p22. Genome Res 2000; 10: 244–57.[Abstract/Free Full Text]
  36. Bookstein R, Bova GS, MacGrogan D, Levy A, Isaacs WB. Tumour-suppressor genes in prostatic oncogenesis: a positional approach. Br J Urol 1997; 79 (Suppl 1): 28–36.
  37. Kerangueven F, Noguchi T, Coulier F, et al. Genome-wide search for loss of heterozygosity shows extensive genetic diversity of human breast carcinomas. Cancer Res 1997; 57: 5469–74.[Abstract/Free Full Text]
  38. Anbazhagan R, Fujii H, Gabrielson E. Allelic loss of chromosomal arm 8p in breast cancer progression. Am J Pathol 1998; 152: 815–9.[Abstract]
  39. Yuan B-Z, Zhou X, Durkin ME, et al. DLC-1 gene inhibits human breast cancer cell growth and in vivo tumorigenicity. Oncogene 2003; 22: 445–50.[CrossRef][Medline]
  40. Hamaguchi M, Meth JL, von Klitzing C, et al. DBC2, a candidate for a tumor suppressor gene involved in breast cancer. Proc Natl Acad Sci U S A 2002; 99: 13647–52.[Abstract/Free Full Text]
  41. Zhou XZ, Lu KP. The Pin2/TRF1-interacting protein PinX1 is a potent telomerase inhibitor. Cell 2001; 107: 347–59.[CrossRef][Medline]
  42. MacGrogan D, Levy A, Bova GS, Isaacs WB, Bookstein R. Structure and methylation-associated silencing of a gene within a homozygously deleted region of human chromosome band 8p22. Genomics 1996; 35: 55–65.[CrossRef][Medline]
  43. Cabeza-Arvelaiz Y, Sepulveda JL, Lebovitz RM, Thompson TC, Chinault AC. Functional identification of LZTS1 as a candidate prostate tumor suppressor gene on human chromosome 8p22. Oncogene 2001; 20: 4169–79.[CrossRef][Medline]
  44. Fujiwara Y, Ohata H, Kuroki T, et al. Isolation of a candidate tumor suppressor gene on chromosome 8p21.3-p22 that is homologous to an extracellular domain of the PDGF receptor beta gene. Oncogene 1995; 10: 891–5.[Medline]
  45. Karnik P, Paris M, Williams BRG, Casey G, Crowe J, Chen P. Two distinct tumor suppressor loci within chromosome 11p15 implicated in breast cancer progression and metastasis. Hum Mol Genet 1998; 7: 895–903.[Abstract/Free Full Text]
  46. Minobe K, Onda M, Iida A, et al. Allelic loss on chromosome 9q is associated with lymph node metastasis of primary breast cancer. Jpn J Cancer Res 1998; 89: 916–22.[Medline]
  47. Tsukamoto K, Ito N, Yoshimoto M, et al. Allelic loss on chromosome 1p is associated with progression and lymph node metastasis of primary breast carcinoma. Cancer 1998; 82: 317–22.[CrossRef][Medline]
  48. Zucali R, Mariani L, Marubini E, et al. Early breast cancer: evaluation of the prognostic role of the site of the primary tumor. J Clin Oncol 1998; 16: 1363–6.[Abstract/Free Full Text]
  49. Fredriksson I, Liljegren G, Arnesson L-G, et al. Local recurrence in the breast after conservative surgery–a study of prognosis and prognostic factors in 391 women. Eur J Cancer 2002; 38: 1860–70.
  50. Lininger RA, Fujii H, Man YG, Gabrielson E, Tavassoli FA. Comparison of loss of heterozygosity in primary and recurrent ductal carcinoma in situ of the breast. Mod Pathol 1998; 11: 1151–9.[Medline]
  51. Regitnig P, Moser R, Thalhammer M, et al. Microsatellite analysis of breast carcinoma and corresponding local recurrences. J Pathol 2002; 198: 190–7.[CrossRef][Medline]
  52. Partridge M, Pateromichelakis S, Phillips E, Emilion GG, A’Hern RP, Langdon JD. A case-control study confirms that microsatellite assay can identify patients at risk of developing oral squamous cell carcinoma within a field of cancerization. Cancer Res 2000; 60: 3893–8.[Abstract/Free Full Text]
  53. Rosin MP, Cheng X, Poh C, et al. Use of allelic loss to predict malignant risk for low-grade oral epithelial dysplasia. Clin Cancer Res 2000; 6: 357–62.[Abstract/Free Full Text]
  54. Gupta D, Nath M, Layfield LJ. Utility of four-quadrant random sections in mastectomy specimens. Breast J 2003; 9: 307–11.[CrossRef][Medline]



This article has been cited by other articles:


Home page
 Ann. Surg. Oncol.Home page
R. E. Ellsworth, D. L. Ellsworth, B. Love, H. L. Patney, L. R. Hoffman, J. Kane, J. A. Hooke, and C. D. Shriver
Correlation of Levels and Patterns of Genomic Instability With Histological Grading of DCIS
Ann. Surg. Oncol., November 1, 2007; 14(11): 3070 - 3077.
[Abstract] [Full Text] [PDF]

Home page
 J. Mol. Diagn.Home page
C. M. Heaphy, W. C. Hines, K. S. Butler, C. M. Haaland, G. Heywood, E. G. Fischer, M. Bisoffi, and J. K. Griffith
Assessment of the Frequency of Allelic Imbalance in Human Tissue Using a Multiplex Polymerase Chain Reaction System
J. Mol. Diagn., April 1, 2007; 9(2): 266 - 271.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
P. S. Yan, C. Venkataramu, A. Ibrahim, J. C. Liu, R. Z. Shen, N. M. Diaz, B. Centeno, F. Weber, Y.-W. Leu, C. L. Shapiro, et al.
Mapping Geographic Zones of Cancer Risk with Epigenetic Biomarkers in Normal Breast Tissue.
Clin. Cancer Res., November 15, 2006; 12(22): 6626 - 6636.
[Abstract] [Full Text] [PDF]

Home page
 Ann. Surg. Oncol.Home page
R. E. Ellsworth, D. L. Ellsworth, B. Deyarmin, L. R. Hoffman, B. Love, J. A. Hooke, and C. D. Shriver
Timing of Critical Genetic Changes in Human Breast Disease
Ann. Surg. Oncol., December 1, 2005; 12(12): 1054 - 1060.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ellsworth, D. L.
Right arrow Articles by Shriver, C. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ellsworth, D. L.
Right arrow Articles by Shriver, C. D.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS