Breast Tissue In Patients Research Articles

P53 alteration in morphologically normal/benign breast tissue in patients with triple-negative high-grade breast carcinomas: breast p53 signature?

p53 alterations have been identified in approximately 23% of breast carcinomas, particularly in hormone receptor-negative high-grade carcinomas. It is considered to be an early event in breast carcinogenesis. Nevertheless, the putative precursor lesion of high-grade breast carcinoma remains elusive. Breast excision specimens from 93 triple-negative high-grade invasive ductal carcinomas, 48 estrogen receptor (ER)-positive/progesterone receptor-positive/Her2-negative non-high-grade invasive ductal carcinomas, and 50 mammoplasty breasts were selected. At least 2 tissue blocks with tumor and adjacent benign tissue were sectioned and subjected to immunohistochemistry staining for p53. TP53 gene sequencing was performed on select tumors. Further immunohistochemistry staining for ER and Ki-67 was performed on consecutive sections of tissue with p53-positive normal/benign cells. Of the 93 high-grade carcinomas, 51 (55%) were positive for p53 alteration, whereas only 3 (6.25%) of the 48 non-high-grade carcinomas were p53 altered. Focal p53 positivity in adjacent normal/benign breast tissue was identified in 19 cases, and 18 of them also had p53 alteration in their carcinomas. Only 1 case had focal p53 staining in normal/benign tissue, but the tumor was negative for p53 alteration. No p53 staining positivity was identified in the mammoplasty specimens. The p53-stained normal/benign cells were ER negative and did not show an increase in the Ki-67 labeling index. These findings indicate that the p53 staining positivity in normal/benign breast tissue is not a random event. It could be considered as the "p53 signature" in breast and serve as an indicator for future potential risk of p53-positive high-grade breast carcinoma.

High levels of sphingolipids in human breast cancer

BackgroundSphingolipids, including sphingosine-1-phosphate (S1P) and ceramide, have emerged as key regulatory molecules that control various aspects of cell growth and proliferation in cancer. Although important roles of sphingolipids in breast cancer progression have been reported in experimental models, their roles in human patients have yet to be determined. The aims of this study were to determine the levels of sphingolipids including S1P, ceramides, and other sphingolipids, in breast cancer and normal breast tissue and to compare the difference in levels of each sphingolipid between the two tissues. Materials and methodsTumor and noncancerous breast tissue were obtained from 12 patients with breast cancer. Sphingolipids including S1P, ceramides, and their metabolites of sphingosine, sphingomyelin, and monohexosylceramide were measured by liquid chromatography–electrospray ionization–tandem mass spectrometry. ResultsThe levels of S1P, ceramides, and other sphingolipids in the tumor were significantly higher than those in normal breast tissue. There was a relatively strong correlation in the levels of S1P between the tumor and those of normal breast tissue from the same person. On the other hand, there was no correlation in the levels of most of the ceramide species between the tumor and those of normal breast tissue from the same person. ConclusionsTo our knowledge, this is the first study to reveal that levels of sphingolipids in cancer tissue are generally higher than those of normal breast tissue in patients with breast cancer. The correlation of S1P levels in these tissues implicates the role of S1P in interaction between cancer and the tumor microenvironment.

Abstract P1-07-08: TP53 alteration in morphologically benign breast tissue in patients with ER/PR negative high grade breast carcinomas: Marker for early detection of high risk patients for high grade carcinomas?

Abstract Background: TP53 alterations have been identified in approximately 20% of breast carcinomas, especially in ER/PR negative high grade carcinomas. It is believed that TP53 alteration is an early event in the carcinogenesis of high grade breast carcinomas. TP53 alteration could even be identified in morphologically "benign" appearing breast tissues. Nevertheless, the precursor lesion of high grade ductal carcinoma in situ and/or invasive carcinoma is not well recognized morphologically. Design: We retrospectively analyzed the TP53 status of 108 cases with high grade (modified Bloom-Richardson grade 3) invasive ductal carcinoma, 51 cases with non-high grade invasive ductal carcinoma, and 50 cases with benign breast from mammoplasty. The high grade carcinomas were all estrogen receptor (ER) and progesterone receptor (PR) negative and 96.3% of them were HER-2 negative. The non-high grade carcinomas were all ER and/or PR positive, 92.2% Her2 negative. Blocks with tumor and/or benign tissue were selected from each case. Immunohistochemical stain for p53 was performed. TP53 gene sequencing was performed on selected tumors with inconclusive p53 staining. Further ER and Ki67 immunohistochemical stains were performed on the blocks with p53 positive benign tissue. Results: Of the 108 high grade carcinomas, 48 (44%) were positive for TP53 alteration. Seventeen of the 48 (35.4%) cases also showed focal p53 staining in the adjacent benign appearing tissue. Only one case was negative for TP53 alteration in tumor but with focal p53 staining in benign tissue. Of the non-high grade carcinomas, 5 (9.8%) were positive for TP53 alteration, one of which had focal p53 staining in adjacent benign breast tissue. No p53 staining positivity was identified in the mammoplasty specimens. The p53 staining positive benign glands were ER negative, and did not show an increase in Ki67 labeling index. The morphology of these cells appeared to be slightly enlarged nuclei, with a mildly increased nuclear/cytoplasm ratio, slightly irregular nuclear contours, open chromatin, and prominent nucleoli, but still within the limits of "benign" glands. Conclusion: TP53 is already altered in "benign" appearing breast glands in patients with high grade breast carcinomas. These glands show the same ER status as their counterpart carcinomas, but remain inactive with low Ki67 labeling index, indicating further genetic changes are needed to trigger the unlimited tumor growth and fully malignant transformation. The special morphology and p53 positivity of the "benign" appearing cells could serve as an indicator for future potential risk of TP53 positive high grade breast carcinoma. Citation Format: Xi Wang, Moritz Stolla, Todd S Laughlin, Paul G Rothberg, Kristin Skinner, David G Hicks. TP53 alteration in morphologically benign breast tissue in patients with ER/PR negative high grade breast carcinomas: Marker for early detection of high risk patients for high grade carcinomas? [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P1-07-08.

Histopathological Features of Non-Neoplastic Breast Parenchyma Do Not Predict BRCA Mutation Status of Patients with Invasive Breast Cancer.

BACKGROUNDSeveral studies have evaluated histologic features of non-neoplastic breast parenchyma in patients with BRCA1/2 mutations, but the results are conflicting. The limited data suggest a much higher prevalence of high-risk precursor lesions in BRCA carriers. Therefore, we designed this study to compare the clinicopathological characteristics of peritumoral benign breast tissue in patients with and without deleterious BRCA mutations.METHODSWomen with breast cancer (BC) who were referred for genetic counseling and underwent BRCA genetic testing in 2010 and 2011 were included in the study.RESULTSOf the six benign histological features analyzed in this study, only stromal fibrosis grade 2/3 was found to be statistically different, with more BRCA noncarriers having stromal fibrosis grade 2/3 than BRCA1/2 carriers (P = 0.04).CONCLUSIONThere is no significant association between mutation risk and the presence of benign histologic features of peritumoral breast parenchyma.

Lobulitis in nonneoplastic breast tissue from breast cancer patients: association with phenotypes that are common in hereditary breast cancer

Lobular inflammation (lobulitis) has been demonstrated in benign breast tissue adjacent to in situ and invasive breast cancers and, more recently, in nonneoplastic tissue from prophylactic mastectomy specimens for hereditary high-risk breast carcinoma. The aim of this study is to investigate the incidence of lobulitis in benign breast tissue of patients with breast cancer and associated clinicopathologic features. We reviewed nonneoplastic breast tissue sections from 334 patients with invasive breast carcinoma to study lobulitis in normal breast tissue and to correlate its presence with clinicopathologic features of the associated tumor. Clinical information (age, menopausal status, and follow-up), tumor characteristics (type, grade, size, lymph node status, stage, estrogen and progesterone receptor, HER2), and survival were recorded. Characteristics of women with and without lobulitis were cross-classified with categories of clinical, pathologic, and histologic characteristics, and differences in distributions were tested in univariate and multivariate analysis. Lobulitis was found in 26 (8%) of 334 patients. The lymphocytic infiltrate was predominantly T-cell type. In a multivariate model, lobulitis in patients with breast cancer was significantly associated with younger age, triple (estrogen receptor, progesterone receptor, HER2)-negative cancers, and medullary phenotypes. Lobulitis in nonneoplastic breast tissue, away from tumor, is associated with clinicopathologic features more commonly seen in hereditary breast cancer.

Abstract 2723: Host response to HMTV

Abstract Work from our laboratory demonstrated the presence of retroviral sequences homologous to the mouse mammary tumor virus (MMTV) in a large proportion of human breast cancers. These observations have been confirmed by others. A 660 bp sequence of the MMTV env gene, with no significant homology to any viral or human sequence reported in the GenBank, has been found in 40% of the U.S. human breast cancers studied. This sequence is expressed in most of the env positive tumors, and is not detected in the normal breast tissue of patients with env positive carcinomas, indicating its exogenous origin. A complete provirus structure with 95% homology to MMTV has been described and designated as human mammary tumor virus (HMTV). Betaretroviral particles from primary cultures of breast cancer cells (MSSM) have been isolated and characterized. Virion RNA is more than 90% homologous to MMTV RNA and to the HMTV proviral DNA previously identified. To find out if these particles were infectious, co-culture experiments between MSSM cells and normal human epithelial breast cells, B and T human lymphocytes and human dendritic cells were performed. The results demonstrated that the recipient human cells became infected with HMTV. However, protein expression was only observed in 10-15% of the infected cells by FACS analysis suggesting the presence of an innate resistance to HMTV in human cells. We have then investigated whether human APOBEC and TRIM proteins are implicated in resistance to HMTV expression. The results indicated that APOBEC G and F, TRIM 5, 21 and 25 are all highly expressed in HMTV cells as measured by quantitative RT-PCR. Additional molecular changes seen in infected cells are disruption of the cytoskeleton and evidence for epithelial-mesenchymal transition (EMT). Since expression of an endogenous retrovirus (HERV-K) env gene has been reported to be upregulated in human breast cancers we have also investigated its expression in HMTV-infected cells, primary MSSM cells and breast tumors by quantitative RT PCR. The results indicated that in all instances where HMTV was present, expression of HERV-K (HML-2) was less than the control counterpart. In conclusion, HMTV is able to infect a variety of cells bringing about striking molecular changes. Whether these changes play a role in tumorigenesis remains to be shown. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2723. doi:10.1158/1538-7445.AM2011-2723

Interleukin 13 expression in the primary breast cancer tumor tissue

The aim of this study was to investigate the presence and the expression levels of the interleukin 13 (I1-13) in the primary breast cancer tumour tissue in relation to the unchanged breast tissue in the same patients and to the breast tissue in the patients with benign breast disease, and to investigate the correlation between the IL-13 expression levels and the pathohistological factors, and between IL-13 expression and estrogens and progesterone receptor status. 50 patients with invasive ductal breast cancer and 20 patients with benign breast diseases were included in this prospective case-control study. The three-step immunohistochemical staining was used for testing the levels of IL-13 expression and hormone receptor status. IL-13 was present in breast cancer tumour tissue, and in the surrounding unchanged tissue in the same patients, and in breast tissue in patients with benign breast disease. The expression of IL-13 was significantly higher in breast cancer tumour compared with surrounding tissue (P < 0.05) of the same, lymph node-positive patients. In addition, IL-13 expression was significantly higher in breast cancer tumour compared with breast tissue in patients with benign breast diseases (P < 0.01). There was significant correlation between IL-13 expression and tumour size in patients with lymph node-negative breast cancer (r = 0.405, P = 0.050). There was no significant correlation between IL-13 expression and the other pathohistological factors, and no significant correlation between IL-13 expression and the lymph node status. Obtained results suggest possible involvement of IL-13 in breast carcinogenesis.

Association between mitochondrial DNA 4,977 bp deletion and NAD(P)H:quinone oxidoreductase 1 C609T polymorphism in human breast tissues

The NAD(P)H:quinone oxidoreductase 1 (NQO1) enzyme is implicated in protection against oxidative stress and carcinogenesis. NQO1 C609T genetic polymorphism was reported to be associated with an increased risk for cancers, including breast cancer. However, there is still lack of evidence whether higher oxidative stress occurs in breast tissues of patients with NQO1 609 C/T and/or T/T genotypes. Mitochondrial DNA (MtDNA) 4,977-bp deletion, the most common mutation of mtDNA, was frequently detected in post-mitotic tissues of aged subjects and associated with oxidative damage. In this study, we detected the mtDNA 4,977-bp deletion in 60 breast cancers and corresponding non-cancerous breast tissues. The incidence of the common 4,977-bp deletion in non-cancerous breast tissues (48.3%) was higher than that in breast cancer (5.0%). Moreover, 63.4% of the breast cancer patients with NQO1 C/T or T/T genotypes had the deletion in their non-cancerous breast tissues. The mtDNA deletion was more frequently detected in breast tissue of NQO1 C/T carriers (65.2%) and T/T carriers (61.1%) as compared with the NQO1 C/C carriers (15.8%, P=0.003). Similar results were observed in the <50 years old (y/o) group (P=0.06) and the > or =50 y/o group (P=0.005). Our findings suggest that mtDNA 4,977-bp deletion associated with NQO1 deficiency is involved in carcinogenesis and progression of breast cancer.

Quantitative MRI assessment of uninvolved breast tissue in patients with locally-advanced breast cancer undergoing neoadjuvant chemotherapy.

Abstract Abstract #4016 Background: We studied the Magnetic Resonance Imaging (MRI) characteristics of breast tissue in patients with invasive cancers to better understand the relationship between tumors and their normal “host” tissue. There is a strong association between tumor morphologic patterns on MRI and both treatment outcome and survival. The objectives of this study were to assess the characteristics of normal host tissue to verify whether they were associated with treatment outcome.&amp;#x2028; Materials and Methods: 42 women with locally-advanced breast cancer underwent MRI exams prior to the start of neoadjuvant adriamycin/cytoxan (AC) chemotherapy (MRI1), following one cycle of AC (MRI2) and at the completion of chemotherapy, prior to surgery (MRI3). Patients mean age was 48.56 years (range 29.7 - 71.5). Median follow-up was 178 weeks. Pathology and radiology reports were available for all patients. Total follow-up was 3 years. We quantified tumor and surrounding normal tissue enhancement. Signal Enhancement Ratio (SER) values have been histopathologically shown to positively correlate with microvessel density within tumor. We lowered the enhancement threshold necessary to detect MRI tumor-enhancing regions, to insure that slow or non-enhancing tissue regions could be analyzed. For all patients, tumor morphologic patterns were categorized according to the degree of tumor containment from 1, denoting well-circumscribed uni-centric masses to 5, corresponding to infiltrative tumors with ill-defined borders. MRI breast tissue composition quantitative measures included 1) volumetric breast density, 2) tumor volume, 3) normal fibroglandular tissue volume and 4) SER levels in tumor and normal surrounding tissue. In order to quantify the uninvolved breast tissue volume, we used a semi-automated technique that performs a segmentation of MRI breast tissue from fat regions without user interaction, therefore avoiding subjective threshold decision issues. Regions in normal tissue surrounding tumors were automatically selected and analyzed at all time points.&amp;#x2028; Results: 10 patients recurred. We found that non-tumor mean SER did not change significantly with one cycle of chemotherapy, however the value of non-tumor mean SER after 1 cycle of chemotherapy was an independent predictor of disease free survival. We also observed a strong association between the initial (pre-treatment) tumor morphologic patterns on MRI with treatment outcome. We found that volume of normal breast tissue at baseline does relate to tumor phenotype (p&amp;lt;0.067), and change in non-tumor breast tissue volume after chemotherapy was also predictive of recurrence (p&amp;lt;0.075).&amp;#x2028; Conclusion: We found that measures of uninvolved breast tissue volume were correlated with tumor phenotypes and were shown to be predictive of recurrence. These results suggest that the host tissue may provide treatment response information. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 4016.

A Technique to Decrease Breast Shape Deformity During Muscle Contraction in Submuscular Augmentation Mammaplasty

The main objective of this study was to decrease breast shape distortion during pectoralis muscle contraction following submuscular augmentation mammaplasty. We followed 348 patients who had retromuscular augmentation mammoplasty: 251 (72.1%) had polyurethane-covered gel-filled, 97 (27.9%) had textured-silicone gel-filled implants. Among the 348, 46 had surgery following the Regnault technique and 302 had surgery by the below-mentioned technique. Periareolar incision, bipartision of breast parenchyma down to the fascia, undermining of breast base from the fascia downward to the inframammary sulcus or a little below it, detaching of muscle off the thoracic cage, disinsertion of abdomino-costal pectoralis attachments. Full thickness incision of pectoralis muscle on a vertical line on the nipple projection for 2-5 inches. Placing of prosthesis. Drainage. Closure. No objective evaluation was used, only clinical judgments by three observers--the surgeon, a nurse and the patient herself. The results showed a definite decrease of the dynamic deformity among patients in whom the author's technical variation was carried out. This technique allows also, for decreasing the upward pushing of the implant during pectoralis muscle contraction and facilitates stretch of the breast tissue in patients with tighter breast envelopes. Despite lack of precise measurements, conclusions drawn from clinical judgments, taken as objectively as possible, suggest that the use of this technique may offer the solution of an otherwise disturbing collateral effect, frequently seen after this operation.

Extra-tumoral breast tissue in breast-cancer patients: variations with a family history of breast cancer. WHO Collaborative Study of Neoplasia and Steroid Contraceptives.

In order to study the relationship between benign breast changes, a family history of breast cancer and breast cancer, extratumoral breast tissue from 1259 breast-cancer patients in the WHO Collaborative Study of Neoplasia and Contraceptives was classified histologically. The occurrence of ductal hyperplasia, ductal atypia, sclerosing adenosis, adenosis, lobular atypia, lactational metaplasia, cysts, apocrine metaplasia, apocrine hyperplasia and atypia, duct ectasia and the epithelial-stromal ratio was evaluated as absent, mild, moderate or marked, along with registration of the quality and number of slides. Information on occurrence of cancer in the family was available for patients' mothers and grandmothers. Logistic-regression analyses showed that the prevalence odds ratios for lactational metaplasia, cysts, duct ectasia and calcification were significantly increased in patients with a family history of breast cancer. Apocrine metaplasia and hyperplasia were not significantly increased. The prevalence rates of ductal atypia (ductal carcinoma in situ and atypical ductal hyperplasia), ductal hyperplasia, sclerosing adenosis, adenosis and high epithelial-stromal ratio did not differ significantly among patients with or without a family history of breast cancer. A family history of other types of cancer did not influence the occurrence of any of the benign components. The findings in the present study are strikingly similar to those in our earlier comparison of extra-tumoral breast tissue in patients from countries with high and low risk of breast cancer. It is reasonable to conclude from this that a history of breast cancer in a woman's mother or grandmother and the factors leading to higher risk of breast cancer in some countries than in others have similar effects on the morphologic evolution of breast cancer through benign and pre-cancerous changes.

Comparison of total cathepsin D in breast tissue of patients with benign and malignant breast disease

Cathepsin D was estimated by immunoradiometric assay in cytosols of breast biopsies from 16 patients with fibrocystic breast disease, 14 patients with fibroadenoma of the breast, 30 patients with ductal carcinoma in situ and seven normal breast tissues from reduction mammoplasties. The results of the groups were compared with one another and with that of a large sample (738) of invasive breast carcinomas from a previous study. The mean cathepsin D concentration in the cytosol was lowest in normal breast tissues, intermediate in ductal carcinomas in situ and adenomas, and highest in invasive carcinomas. Cathepsin D in ‘fibrocystic disease’ covered a wide range; the average was still below the cut off value of 25 pmol/mg protein, but closer to the mean of the invasive carcinomas than that of any other group.

Analysis of silicon levels in capsules of gel and saline breast implants and of penile prostheses.

Although a potential link between silicone-gel breast implants and autoimmune connective tissue disease has been suggested, none has been proven. The potential role of silicone as an immune adjuvant remains very controversial. Currently available techniques do not allow precise measurements of silicone in tissues. However, all compounds containing silicon (including silicone) can be measured accurately. The present study was designed to measure silicon levels in the fibrous capsules of patients with silicone-gel breast implants, saline breast implants, and silicone inflatable penile prostheses. Baseline control silicon levels were obtained from the breast tissue of patients undergoing breast reduction, who had no exposure to breast implants. All silicon measurements were carried out using atomic absorption spectrometry with a graphite furnace. Silicon was measured in a normal heptane extract of silicone from dried tissue. The mean silicon levels in 16 breast tissue control samples from 8 patients undergoing breast reduction varied from 0.025 to 0.742 micrograms/gm with the median mean being 0.0927. The median silicon level in capsules from six patients with saline implants was 7.7 micrograms/gm (range, 1.9-36.6 micrograms/gm). The median silicon level in capsules from five patients with silicone inflatable penile prostheses was 19.5 micrograms/gm (range, 1.9-34.8 micrograms/gm). Although the levels of silicon in capsules of patients with saline breast prostheses and penile implants were higher than in control samples, they were much lower than those from the capsules of the 58 gel implants (median, 9,979 micrograms/gm; range, 371-152,000 micrograms/gm).(ABSTRACT TRUNCATED AT 250 WORDS)

Technical considerations for prophylactic mastectomy in patients at high risk for breast cancer

A study of 5 patients and 10 mastectomy specimens was performed to identify the extent of surgery necessary to completely remove all breast tissue in patients having prophylactic mastectomies. A standard total mastectomy performed for breast cancer was shown to frequently leave breast tissue within the superficial pectoralis major muscle and the lower skin flap. Frozen section analysis of margins was found to be essential to clear the axillary extension of the breast and lower skin flap in particular. The value of more extensive surgery to remove all glandular elements of the breast in the high-risk patient remains to be demonstrated.

Importance of estrogen sulfates in breast cancer

Estrogen sulfates are quantitatively the most important form of circulating estrogens during the menstrual cycle and in the post-menopausal period. Huge quantities of estrone sulfate and estradiol sulfate are found in the breast tissues of patients with mammary carcinoma. It has been demonstrated that different estrogen-3-sulfates (estrone-3-sulfate, estradiol-3-sulfate, estriol-3-sulfate) can provoke important biological responses in different mammary cancer cell lines: there is a significant increase in progesterone receptor. On the other hand, no significant effect was observed with estrogen-17-sulfates. The reason for the biological response of estrogen-3-sulfates is that these sulfates are hydrolyzed, and no sulfatase activity for C17-sulfates is present in these cell lines. [3H]Estrone sulfate is converted in a very high percentage to estradiol (E2) in different hormone-dependent mammary cancer cell lines (MCF-7, R-27, T-47D), but very little or no conversion was found in the hormone-independent mammary cancer cell lines (MDA-MB-231, MDA-MB-436). Different anti-estrogens (tamoxifen and derivatives) and another potent anti-estrogen: ICI 164,384, decrease the concentration of estradiol very significantly after incubation of estrone sulfate with the different hormone-dependent mammary cancer cell lines. No significant effect was observed for the uptake and conversion of estrone sulfate in the hormone-independent mammary cancer cell lines. Progesterone provokes an important decrease in the uptake and in estradiol levels after incubation of [3H]estrone sulfate with the MCF-7 cells. It is concluded that in breast cancer: (1) Estrogen sulfates can play an important role in the biological response of estrogens; (2) Anti-estrogens and progesterone significantly decrease the uptake and estradiol levels in hormone-dependent mammary cancer cell lines; (3) The control of the sulfatase and 17 beta-hydroxysteroid dehydrogenase activities, which are key steps in the formation of estradiol in the breast, can open new possibilities in the treatment of hormone-dependent mammary cancer.