Malignant Breast Tissue Research Articles

LIF spectroscopy of stained malignant breast tissues.

We employ laser induced fluorescence (LIF) spectroscopy to discriminate between normal and cancerous human breast (in-vitro) tissues. LIF signals are usually enhanced by the exogenous agents such as Rhodamine 6G (Rd6G) and Coumarin 7 (C7). Although we observe fluorescence emissions in both fluorophores, Rd6G-stained tissues give notable spectral red shift in practice. The latter is a function of dye concentration embedded in tissues. We find that such red shifts have a strong dependence on the dye concentration in bare, in stained healthy, and in malignant breast tissues, signifying variations in tubular abundances. In fact, the heterogeneity of cancerous tissues is more prominent mainly due to their notable tubular densities- which can provide numerous micro-cavities to house more dye molecules. We show that this can be used to discriminate between the healthy and unhealthy specimens in different biological scaffolds of ordered (healthy) and disordered (cancerous) tissues. It is demonstrated that the quenching process of fluorophore' molecules slows down in the neoplastic tumors according to the micro-partitioning, too.

18F-Fluorocholine PET/CT of Incidental Male Breast Cancer

Breast cancer is approximately 100 times less common among men. Estrogen-related receptor α was detected in human prostate cancer tissue, and androgen receptors are expressed in both normal and malignant breast tissue. Thus, prostate cancer hormonal therapy may increase breast cancer risk. A 67-year-old man with prostate cancer history presented with rising prostate-specific antigen level. F-Fluorocholine PET/CT showed intense uptake in his right breast and moderate uptake in right axillary lymph node. Complementary F-FDG PET/CT demonstrated more intense tracer uptakes in both the right breast and axillary lymph node. Breast histological findings were invasive ductal cancer, with axillary lymph node invasion.

Is HPV virus associated with human breast cancer? Time to re-examine the postulate. Egyptian Study

Background: There is increasing evidence that high-risk human papilloma virus (HPV) is involved in cancers in addition to cervical cancer. Infectious agents are thought to be responsible for approximately 16% of cancers worldwide, however there are mixed reports in the literature regarding the prevalence and potential pathogenicity of viruses in breast cancer. Methods : We screened 30 fresh frozen breast cancer tissue specimens collected immediately postoperative from patients in Ain-Shams University Hospital for the presence of human papilloma virus (HPV) DNA by PCR and Koilocytic changes in the breast cancer tissue. Results: Overall prevalence of HPV in malignant breast tissue was 16.7% In addition, we found that the oncogenic characteristics of HPV associated breast cancer are very similar to HPV-associated cervical cancer. Specifically, that putative koilocytes are present in some HPV associated breast cancers. Conclusion : The above observations indicate a likely causal role for high-risk HPV in human breast cancer and offer the possible role of HPV in breast cancer prognosis. DOI: 10.21276/APALM.2017.1143

Identification of Potential Tumor Markers in Sudanese Breast Cancer Patients Using a Proteomic Approach

Breast cancer (BC) is considered as a major health problem in Sudan, being the most frequent hospital treated malignancy and the commonest cancer affecting Sudanese females (34%). Proteomics provides tools that investigate the precise molecular defect(s) in breast cancer tissue. Moreover, it plays a growingly important role in tumor markers discovery. In this study, a proteomic assay (2D-PAGE) was used to investigate the protein profiles of a panel of 12 Sudanese breast malignant tissues and 12 matched controls. Protein spots of interest were excised manually, and in-gel digested using trypsin. Proteins were identified using mass spectrometry (MALDITOF). Mascot program was used to search protein databases for matching peptides from known proteins. The overall profile was relatively similar, the preliminary results identified three proteins to be differentially expressed, suggesting that they may perform a role in breast neoplasia. Although the number of samples investigated in this study is comparatively small, to allow authoritative conclusions, the study provided three proteins that might be potential tumor markers in Sudanese breast cancer patients requiring further investigations and validation.

Optical Properties of Non-Malignant and Malignant Breast Tissue Determined by Surface Displacement of Laser-Induced Photoacoustic Generation

Optical Properties of Non-Malignant and Malignant Breast Tissue Determined by Surface Displacement of Laser-Induced Photoacoustic Generation

Gene expression profiles of CYP24A1 and CYP27B1 in malignant and normal breast tissues.

Active vitamin D has several antitumor effects, including prodifferentiative, antiproliferative and proapoptotic functions in a number of tissues via its binding to vitaminD receptor. The 24‑hydroxylase (CYP24A1) and 1‑hydroxylase (CYP27B1) enzymes are considered to be pivotal determinants of the local concentration of active vitamin D. The aim of the present study was to investigate the mRNA expression levels of the CYP24A1 and CYP27B1 genes in malignant and normal breast tissues. The tumor and adjacent normal tissue samples of 30patients with breast cancer were acquired from the Iran National Tumor Bank, Imam Hospital (Tehran, Iran). RNA was extracted and, following cDNA synthesis, Taq‑Man quantitative polymerase chain reaction analysis was performed for CYP24A1 and CYP27B1. The results demonstrated that the mRNA expression of CYP27B1 was downregulated in the tumor tissues, compared with the adjacent normal tissues (P<0.01), whereas the mRNA expression of CYP24A1 was significantly upregulated in the tumor tissues (P<0.01). This major difference revealed that the normal breast tissues transcriptionally expressed CYP24A1 slightly. These results are suggestive of dysregulation of the vitamin D signaling and metabolic pathways during tumorigenesis in breast cancer. Local alterations in the anabolism and catabolism of active vitaminD in breast cancer by the CYP27B1 and CYP24A1 may impair its anticancer functions.

The Expression of Cohesin Subunit SA2 Predicts Breast Cancer Survival

Cohesin is one of the main regulators of sister chromatid separation during the metaphase/anaphase transition. It is a multiprotein complex consisting of 4 core subunits, one of those being the SA2 subunit. SA2 plays the final role in dismantling the cohesion complex from the sister chromatids and also functions in DNA double-strand break repair and gene regulation. There is increasing evidence regarding the involvement of both overexpression and underexpression of cohesin in cancer. Here, we present expression patterns of SA2 in different types of human breast tissue, and the prognostic analysis in the material from breast cancer patients with long-term follow-up. SA2 immunoexpression was evaluated in benign, precancerous, and malignant breast tissue, and was classified into low-intensity or high-intensity groups. The DNA content was determined by image cytometry on breast cancer cell imprints. Prognostic analyses were based on 445 breast cancer patients with upto 20 years' follow-up. SA2 immunoexpression was equally high in both benign and precancerous breast tissue. Instead, 72% of the invasive breast cancers showed deficient SA2 expression. These patients were also associated with an unfavorable outcome as indicated by a 1.6-fold risk of breast cancer death (P=0.0208). The majority (75%) of the patients with low SA2 expression were alive 6.0 years after the diagnosis, whereas the majority of the patients with high SA2 expression survived 17.6 years after the diagnosis. No statistically significant association could be detected between SA2 immunoexpression and DNA aneuploidy. Our results and previous literature indicate that decreased SA2 immunoexpression is associated with malignant breast disease and a particularly unfavorable course of disease.

Sprouty4 mRNA variants and protein expressions in breast and lung-derived cells.

Sprouty proteins are modulators of mitogen-induced signalling processes and are therefore hypothesized to affect malignant diseases. As a member of the Sprouty family, Sprouty4 has been previously shown to function as a tumour suppressor in lung and breast cancer. The present study analysed the expression of two known Sprouty4 splice variants in cells established from malignant and normal lung and breast tissues using semi-quantitative reverse transcription-polymerase chain reaction and immunoblotting. The results indicated that the expression of the two messenger RNA (mRNA) variants was reduced in the cells derived from malignant tissue in comparison to the normal counterparts. Although the expression of the two splice variants were associated in both tissue types, on average, the relative expression of the longer variant was slightly increased in malignant cells compared with normal tissues. Notably, the protein levels reflected the expression observed at the mRNA level only in breast-derived cells. Contrarily, with regards to the measured mRNA levels, Sprouty4 protein was disproportional augmented in lung cells known to harbour the mutated K-Ras gene.

Characterization of CD44+ALDH1+Ki-67- Cells in Non-malignant and Neoplastic Lesions of the Breast.

Cancer stem cells are tumor cells that present self-renewal, clonal tumor initiation capacity and clonal long-term repopulation potential. We have previously demonstrated that the co-expression of the breast cancer stem cell (BCSC) markers hyaluronan receptor (CD44) and aldehyde dehydrogenase-1 (ALDH1) in ductal carcinomas in situ could be determinant for disease progression. Combining these established BCSC markers with Ki-67 to evaluate quiescence we sought to identify, evaluate the distribution and estimate the mean percentages of CD44(+)ALDH1(+)Ki-67(-) breast cells. Triple-immunohistochemistry for CD44, ALDH1 and Ki-67 was applied in a series of 16 normal, 54 non-malignant and 155 malignant breast tissues. Clinical relevance was inferred by associations with markers of breast cancer behavior, progression and survival. The mean percentages of cells with this phenotype increased significantly from non-malignant lesions to high-grade ductal carcinomas in situ, decreasing in invasive ductal carcinomas, as also evidenced by an inverse correlation with histological grade and tumor size. The mean percentage of CD44(+)ALDH1(+)Ki-67(-) cells was also significantly higher in women who developed distant metastasis and died due to breast cancer, and a significant association with human epidermal growth factor type 2 (HER2) negativity was observed. Our novel findings indicate that CD44(+)ALDH1(+)Ki-67(-) tumor cells may favor distant metastasis and can predict overall survival in patients with ductal carcinomas of the breast. More importantly, quiescence may have a crucial role for tumor progression, treatment resistance and metastatic ability of BCSCs.

Abstract 3934: Analysis of PKC-ζ protein and mRNA levels in normal and malignant breast tissue

Abstract It is estimated that in 2015 breast cancer will be the second leading cause of cancer death in women. For this purpose, biochemical markers for breast cancer have been investigated, to assist in early detection and more accurate diagnoses. In breast cancer tissue, the atypical isozyme of protein kinase C zeta, PKC-ζ, has been a topic in research; It has been shown that an overexpression of this protein may be indicative of developing carcinogenesis and contributes to proliferation through the NF kappa B pathway, which is a stress-regulated switch for cell survival. In this investigation, the expression of PKC-ζ was analyzed in normal and malignant female human breast tissue samples by Western blot, immunoprecipitation and PCR. In the preliminary results, the malignant breast tissue samples illustrated a significant expression of PKC ζ when compared to the expression of PKC-ζ in normal breast tissue samples. The same tissues were also processed for total RNA isolation which was followed by cDNA synthesis and Real Time PCR. The level of PKC-ζ mRNA was tested and no overexpression was observed in either malignant or normal breast tissue samples. While protein studies may suggest that PKC-ζ could be considered a biomarker for breast cancer, the same cannot be said about mRNA levels. The overexpression of PKC-ζ protein level and the normal PKC-ζ mRNA level are indicators of possible miRNA activity that may regulate translation in malignant tissues but not the normal. Citation Format: Christopher Apostolatos, Tracess Smalley, Mildred A. Duncan. Analysis of PKC-ζ protein and mRNA levels in normal and malignant breast tissue. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3934.

Detection of breast cancer using electromagnetic techniques: A review

Breast cancer is the most well-known malignancy of non-skin and the second leading reason of cancer death among the women in the world after the lung cancer. In this review, the development of the microwave imaging system is described for breast cancer detection depending on the ultra-wideband imaging system, which is a promising emerging technology that uses the dielectric contrast between malignant, benignant and normal fibro glandular breast tissues at the microwave frequen- cies. Some breast cancer detection techniques such as Magnetic Resonance Imaging (MRI), Mammography, Ultra-Wide Band (UWB), Ultrasound and their performances have also been reviewed. There are some restrictions of the existing UWB imaging systems such as they have no ability to detect the breast tumor at the beginning period, having no 3D pictorial representation of tumor size in all systems and a multiple array antenna in respect of complex system. This encourages the improvement of the extended signal processing techniques for ultra-wideband imaging systems and initiates a more appealing imaging scenario. The limitations and running challenges of the microwave imaging system have been discussed completely with some feasible suggestions. Confidently, this thorough survey has been done here to overcome this limitations of the existing and proposed systems.

BREAST CONSERVATIVE SURGERY. A SURGICAL PROCEDURE WHICH COMBINES ONCOLOGICAL SAFETY AND ESTHETIC EFFECT

In the last twenty years, the surgical treatment of breast cancer (BC) has known a substantial progress, evolving from radical mastectomy – a mutilating procedure commonly performed in until 1970s – to breast conservative surgery (BCS) which allows the removal of the entire breast parenchyma saving the skin envelope of the mammary gland, and sentinel lymph node biopsy that helped reducing the complication rate associated with radical axillary dissection. As it has been proved that BCS and radical mastectomy are associated with similar survival rates, BCS or “skin sparing mastectomy” is nowadays considered to be an oncologically safe procedure in locally advanced BC if the surgical resection criteria is respected: negative surgical margins of resection with an entire removal of all the malignant breast tissue and the maintain of aesthetic result. Moreover, owing to the development of novel oncoplastic techniques and the fact that performing BCS allows the preservation of the skin and sometimes the nipple-areola complex (NAC), BCS techniques such as the skin – sparing mastectomy and nipple-areola complex mastectomy are currently considered mainstay treatment of early stage BC without compromising the oncological safety of mastectomy.

Breast Cancer Detection by Determination of Optical Properties of Non-Malignant and Malignant Breast Tissues

Breast cancer being one of the most frequent form of cancer, is the leading cause of cancer deaths in women worldwide. Early detection can greatly improve a womans chances for survival. In this paper we introduce an economic and accurate optical system for breast cancer detection. We have investigated the optical properties of human breast tissues using the He:Ne laser and optical spectroscopy. The materials investigated included non-malignant and malignant breast tissue, which almost have similar grain sizes and thicknesses. The informative features of calculated optical properties of breast tissue at 633 nm allow one to discriminate between normal and malignant sample also Monte Carlo simulation are used to describe the propagation of light . An experimental scheme is applied for detection and imaging the reflected light from the sample to extract information of infected tissue. Calculations of the results demonstrate that the absorption coefficient for cancerous tissue is 10.07 cm -1 relative to 33.66 cm -1 for non-malignant (normal) breast tissue and the laser reflectance is 770.5 cm -1 for cancerous tissue relative to 261.43 cm -1 for normal tissue, which can be used for diagnostic purposes.

Expression and clinical significance of miRNAs that may be associated with the FHIT gene in breast cancer

The dysregulation of miRNA expression has frequently been observed in breast cancer. Therefore, we investigated the expression profile of miRNAs that may be associated with expression of the FHIT gene in breast cancer and assessed their clinicopathological significance. The expression levels of miR-143, miR-663a, miR-668, miR-922 and FHIT were analyzed in normal and malignant breast tissues from 65 patients with breast cancer. We studied the correlation between the expression of miR-143, miR-663a, miR-668, miR-922 and FHIT and the clinicopathological features presented by the patients. The expression levels of the miRNAs and FHIT were downregulated in breast cancer tissue. The expression levels of miR-143, miR-663a and miR-668 were significantly reduced in FHIT downregulated tumors. miR-668 expression was also significantly altered relative to FHIT down- and up- regulated tumor tissues. Reduced miR-663a expression was statistically associated with high-grade ER/PR (+) status, benign reactive hyperplasia, lymph-node metastasis, in-situ component >25% and Ki 67>15% compared with non-tumor tissues. Additionally, reduced miR-668 expression was significantly different between tumors with and without lymph-node metastasis. miR-668 may play an important role in breast cancer development and progression by regulating the expression of FHIT. Furthermore, miR-668 and miR-663a may be potential prognostic biomarkers for breast cancer.

Potential of Diffusion-Weighted Imaging in the Characterization of Malignant, Benign, and Healthy Breast Tissues and Molecular Subtypes of Breast Cancer.

The role of apparent diffusion coefficient (ADC) in the diagnosis of breast cancer and its association with molecular biomarkers was investigated in 259 patients with breast cancer, 67 with benign pathology, and 54 healthy volunteers using diffusion-weighted imaging (DWI) at 1.5 T. In 59 breast cancer patients, dynamic contrast-enhanced MRI (DCEMRI) was also acquired. Mean ADC of malignant lesions was significantly lower (1.02 ± 0.17 × 10−3 mm2/s) compared to benign (1.57 ± 0.26 × 10−3 mm2/s) and healthy (1.78 ± 0.13 × 10−3 mm2/s) breast tissues. A cutoff ADC value of 1.23 × 10−3 mm2/s (sensitivity 92.5%; specificity 91.1%; area under the curve 0.96) to differentiate malignant from benign diseases was arrived by receiver operating curve analysis. In 10/59 breast cancer patients, indeterminate DCE curve was seen, while their ADC value was indicative of malignancy, implying the potential of the addition of DWI in increasing the specificity of DCEMRI data. Further, the association of ADC with tumor volume, stage, hormonal receptors [estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor (HER2)], and menopausal status was investigated. A significant difference was seen in tumor volume between breast cancer patients of stages IIA and IIIA, IIB and IIIA, and IIB and III (B + C), respectively (P < 0.05). Patients with early breast cancer (n = 52) had significantly lower ADC and tumor volume than those with locally advanced breast cancer (n = 207). No association was found in ADC and tumor volume with the menopausal status. Breast cancers with ER−, PR−, and triple-negative (TN) status showed a significantly larger tumor volume compared to ER+, PR+, and non-triple-negative (nTN) cancers, respectively. Also, TN tumors showed a significantly higher ADC compared to ER+, PR+, and nTN cancers. Patients with ER− and TN cancers were younger than those with ER+ and nTN cancers. The present study demonstrated that ADC may increase the diagnostic specificity of DCEMRI and be useful for treatment management in clinical setting. Additionally, it provides an insight into characterization of molecular types of breast cancer and may serve as an indicator of metabolic reprograming underlying tumor proliferation.

Rapid staining and imaging of subnuclear features to differentiate between malignant and benign breast tissues at a point-of-care setting.

Histopathology is the clinical standard for tissue diagnosis; however, it requires tissue processing, laboratory personnel and infrastructure, and a highly trained pathologist to diagnose the tissue. Optical microscopy can provide real-time diagnosis, which could be used to inform the management of breast cancer. The goal of this work is to obtain images of tissue morphology through fluorescence microscopy and vital fluorescent stains and todevelop a strategy to segment and quantify breast tissue features in orderto enable automated tissue diagnosis. We combined acriflavine staining, fluorescence microscopy, and a technique called sparse component analysis to segment nuclei and nucleoli, which are collectively referred to as acriflavine positive features (APFs). A series of variables, which included the density, area fraction, diameter, and spacing of APFs, were quantified from images taken from clinical core needle breast biopsies and used to create a multivariate classification model. The model was developed using a training data set and validated using an independent testing data set. The top performingclassification model included the density and area fraction of smaller APFs (those less than 7µm in diameter, which likely correspond to stained nucleoli).When applied to the independent testing set composed of 25 biopsy panels, the model achieved a sensitivity of 82%, a specificity of 79%, and an overall accuracy of 80%. These results indicate that our quantitative microscopy toolbox is a potentially viable approach for detecting the presence of malignancy in clinical core needle breast biopsies.

Head-to-head comparison of the impact of Aurora A, Aurora B, Repp86, CDK1, CDK2 and Ki67 expression in two of the most relevant gynaecological tumor entities.

The dysregulation of cell cycle kinases plays a crucial role in carcinogenesis and the expression of various kinases has been attributed to aggressive tumor growth and an unfavourable prognosis in oncological patients. We, therefore, aimed to evaluate the expression of Ki67 among five additional cell cycle kinases in a collective of mammary and ovarian tumor specimens and to find a correlation with clinicopathological parameters. 76 mammary and 93 ovarian benign and malignant tumor samples were immunohistochemically stained and evaluated for the expression of Aurora A and B, Repp86, CDK1 and 2 (only breast specimens) and Ki67. The expression patterns of these cell cycle kinases were matched with retrospectively collected clinicopathological parameters. All examined cell cycle kinases accurately discriminated benign from malignant breast and ovarian tissues. In breast cancer, Aurora A and B-, Repp86-, CDK2- and Ki67-expression was inversely associated with ER expression. No correlation with the HER2-status was found in our collective. Importantly, we found a significant correlation between the expression of Aurora A and CDK1 and axillary lymph node metastasis in breast cancer. Furthermore, a shortened disease free survival (DFS) upon expression of Aurora B and CDK2 was shown in breast cancer patients. None of the cell cycle kinases was associated with predictive or prognostic factors in epithelial ovarian cancer. The prognostic value of the expression of Ki67 is overtrumped by alternative cell cycle kinases when it comes to prediction of axillary tumor spread and a shortened DFS, which might allow a further risk stratification in breast cancer patients.

Spectrally encoded confocal microscopy for diagnosing breast cancer in excision and margin specimens

A large percentage of breast cancer patients treated with breast conserving surgery need to undergo multiple surgeries due to positive margins found during post-operative margin assessment. Carcinomas could be removed completely during the initial surgery and additional surgery avoided if positive margins can be determined intraoperatively. Spectrally encoded confocal microscopy (SECM) is a high-speed reflectance confocal microscopy technology that has a potential to rapidly image the entire surgical margin at subcellular resolution and accurately determine margin status intraoperatively. In this study, in order to test the feasibility of using SECM for intraoperative margin assessment, we have evaluated the diagnostic accuracy of SECM for detecting various types of breast cancers. Forty-six surgically removed breast specimens were imaged with an SECM system. Side-by-side comparison between SECM and histologic images showed that SECM images can visualize key histomorphologic patterns of normal/benign and malignant breast tissues. Small (500 μm × 500 μm) spatially registered SECM and histologic images (n=124 for each) were diagnosed independently by three pathologists with expertise in breast pathology. Diagnostic accuracy of SECM for determining malignant tissues was high, average sensitivity of 0.91, specificity of 0.93, positive predictive value of 0.95, and negative predictive value of 0.87. Intra-observer agreement and inter-observer agreement for SECM were also high, 0.87 and 0.84, respectively. Results from this study suggest that SECM may be developed into an intraoperative margin assessment tool for guiding breast cancer excisions.

MiR-449a promotes breast cancer progression by targeting CRIP2.

The identification of prognostic biomarkers and their underlying mechanisms of action remain of great interest in breast cancer biology. Using global miRNA profiling of 71 lymph node-negative invasive ductal breast cancers and 5 normal mammary epithelial tissues, we identified miR-449a to be highly overexpressed in the malignant breast tissue. Its expression was significantly associated with increased incidence of patient relapse, decreased overall survival, and decreased disease-free survival. In vitro, miR-449a promoted breast cancer cell proliferation, clonogenic survival, migration, and invasion. By utilizing a tri-modal in silico approach for target identification, Cysteine-Rich Protein 2 (CRIP2; a transcription factor) was identified as a direct target of miR-449a, corroborated using qRT-PCR, Western blot, and luciferase reporter assays. MDA-MB-231 cells stably transfected with CRIP2 demonstrated a significant reduction in cell viability, migration, and invasion, as well as decreased tumor growth and angiogenesis in mouse xenograft models. Our data revealed that overexpression of miR-449a suppresses CRIP2, which then affects the tumor vasculature, likely via NF-κB/p65 complex-mediated transcription of VEGF. These finding define an oncogenic function of miR-449a in human breast cancer, and highlight the importance of this pathway in driving aggressive behaviour.

Measurement of linear attenuation coefficients of normal and malignant breast tissues using synchrotron radiation

The linear attenuation coefficients of normal and malignant breast tissues have been measured in the energy range of 8–18 keV using Indus‐2 synchrotron source. The intensity of the incident as well as transmitted X‐ray photons in normal and malignant breast tissues is measured with AXUV 100GX silicon photodiode detector. Using these intensities, the linear attenuation coefficients of normal and malignant breast tissues have been measured. The measured linear attenuation coefficient values of cancer tissue are found to be higher than that of normal tissue throughout measured energy region. Copyright © 2016 John Wiley &amp; Sons, Ltd.