Plasma Of Breast Cancer Patients Research Articles

Effect of Radiotherapy on Antitumor Immune Response in Breast Cancer Patients

Background: Radiotherapy induces tumor cell death through DNA damage. Direct relationship between radiation therapy and the immune system is responsible for many of the antitumor effects of radiotherapy. The effects of radiation therapy extend beyond direct cytotoxicity on tumor cells to additional systemic antitumor effects. The objective of this study was to evaluate the effect of radiotherapy on the circulating levels of tumor necrosis factor-α (TNF-α), Interferon-gamma (IFN-γ), CD8+ cytotoxic T lymphocytes and CD56+ natural killer cells in breast cancer patients. Methods: This study included 90 women divided into two main groups: Group I: 45 patients with stage II-III invasive breast carcinoma, group II: 45 healthy women as a control group. All studied patients received adjuvant radiotherapy after surgery. Enzyme-linked immunosorbent assay provided the measurement of TNF-α and INF-γ. Flow cytometry assessed the levels of CD8+ cytotoxic T lymphocytes and CD56+ natural killer cells. Results: A significant reduction occurred in the percent of CD8+ and CD56+ cells in breast cancer patients after radiotherapy. There was a significant increase in both TNF-α and INF-γ in the plasma of breast cancer patients after radiotherapy. Conclusions: TNF-α and INF-γ cytokines significantly increased while the percent of CD56+ and CD8+ cells significantly decreased following exposure to radiotherapy. Such immune modulatory potential and increase in the knowledge of radiation induced out of field and systemic effects, foresee a rapid progress in the development and clinical application of new combined radiotherapeutic and immunotherapeutic approaches.

Tracking Drug-Induced Epithelial-Mesenchymal Transition in Breast Cancer by a Microfluidic Surface-Enhanced Raman Spectroscopy Immunoassay.

Epithelial-mesenchymal transition (EMT) is a primary mechanism for cancer metastasis. Detecting the activation of EMT can potentially convey signs of metastasis to guide treatment management and improve patient survival. One of the classic signatures of EMT is characterized by dynamic changes in cellular expression levels of E-cadherin and N-cadherin, whose soluble active fragments have recently been reported to be biomarkers for cancer diagnosis and prognosis. Herein, a microfluidic immunoassay (termed "SERS immunoassay") based on sensitive and simultaneous detection of soluble E-cadherin (sE-cadherin) and soluble N-cadherin (sN-cadherin) for EMT monitoring in patients' plasma is presented. The SERS immunoassay integrates in situ nanomixing and surface-enhanced Raman scattering readout to enable accurate detection of sE-cadherin and sN-cadherin from as low as 10 cells mL-1 . This assay enables tracking of a concurrent decrease in sE-cadherin and increase in sN-cadherin in breast cancer cells undergoing drug-induced mesenchymal transformation. The clinical potential of the SERS immunoassay is further demonstrated by successful detection of sE-cadherin and sN-cadherin in metastatic stage IV breast cancer patient plasma samples. The SERS immunoassay can potentially sense the activation of EMT to provide early indications of cancer invasions or metastasis.

MMP-2 levels evaluation and their relationship with breast cancer progression.

Matrix metalloproteinases (MMPs) play significant part in the tumor development, metastasis and invasion by proteolytic degradation of basement membrane and extracellular matrix. Hence MMPs can serve as good biomarkers for carcinoma breast particularly MMP-2 in our population. Objectives: To measure the concentrations of MMP-2 in plasma of breast cancer patients and establishing their correlation with stage and grade of the disease. Study Design: Comparative cross sectional. Setting: Surgery department Madinah Teaching Hospital (MTH) and the Pathology department, The University of Faisalabad (TUF), Faisalabad. Period: 1st August, 2017 to 31st December, 2017. Material & Methods: Only females with diagnosed breast cancer disease along with staging and grading were included in the study. Females with incomplete reports, on chemotherapy or radiotherapy or having other diseases like oral squamous cell carcinoma, rheumatoid arthritis etc were excluded. ELISA kit was applied to measure the concentrations of MMP-2 in plasma of the participants. OD values were calculated at 450 nm by ELISA reader placed at Post Graduate Research Laboratory, The University of Faisalabad, Faisalabad. Data was gathered and results were finalized by using SPSS 22 for statistical assessment. Results: Age of 64(72%) breast cancer patients was within range of 30-76yrs and age for 25(28%) healthy females as controls range was 18-70yrs. Difference between ages of cases and controls using t-test was statistically highly significant. Ninety five percent cases showed invasive ductal carcinoma (IDC) as histological type. Range of MMP-2 levels in cases was 80-690 ng/mL with mean 451.02 while range of MMP-2 levels in controls was 65-830ng/mL with mean 329.72 showing statistically significant difference. Relationship of different groups of MMP-2 levels in cases with stages of breast cancer was established using chi-square test which was statistically significant, stage IIB (37.5%) and IIIA (34.4%) tumors showed higher values of MMP-2. Relationship between MMP-2 levels, grade, tumor size and node invasion was nonsignificant statistically. Conclusion: MMP-2 levels were raised in cases in comparison to healthy controls. Age of majority patients was > 45 yrs while the invasive ductal carcinoma was the main histological type. MMP-2 levels were associated with stage of the disease.

The expression pattern of miR-16, 21, 145, and 375 in plasma of breast cancer patients at different clinical stages

The expression pattern of miR-16, 21, 145, and 375 in plasma of breast cancer patients at different clinical stages

A guide to hyaluronan and related enzymes in breast cancer: biological significance and diagnostic value.

Hyaluronan (HA) is a unique nonsulfated glycosaminoglycan that contributes to breast cancer cells growth and functional properties, including cell migration, invasion, adhesion, as well as tumor-associated angiogenesis in different stages of breast cancer progression and especially metastasis. Latest data show that the levels of HA and/or low molecular mass HA in blood serum and plasma of breast cancer patients may be a useful biomarker for breast cancer prognosis, differential diagnosis, and patients' treatment monitoring. Therefore, the qualitative and quantitative determination of HA in biological samples is an emerging area of research. This review gathers, categorizes, and sums up all the currently used methodologies to analyze HA and HA-related enzymes. The advantages, disadvantages, limitations in use, and the information they provide, are critically considered and discussed. Moreover, emphasis is given to the significance of HA determination in breast cancer, as well as of its related enzymes, for diagnosis and prognosis of this type of cancer.

Monitoring Breast Cancer Response to Treatment Using Stokes Shift Spectroscopy of Blood Plasma.

With the emerging trend of personalized cancer treatment, there is a need to develop noninvasive/minimally invasive techniques for treatment monitoring. In this regard, in this work fluorescence analysis of blood plasma of breast cancer patients has been used for the evaluation of response to treatment. This approach delivers information not only about the change in biochemical constituents but also about the altered metabolic pathway. Spectral deconvolution method is employed to compute the fluorescence intensity, peak wavelength, and full-width half maxima for different endogenous fluorophores. The fluorescence measurements were made on blood plasma collected from 10 normal subjects, 10 pre-treated cancer patients, and 10 post-treated patients. Besides, variations in relative concentration of tryptophan, collagen, NADH, and FAD, peak shifts and broadening of peaks are observed for tryptophan, NADH, and FAD, in blood plasma of pre-treated cancer patients indicating both biochemical and microenvironmental changes at cellular level. Further, the spectral profile of blood plasma of post-treated patients found to be similar to blood plasma of normal subjects. Linear discriminant analysis showed that pre-treated and post-treated breast cancer is discriminated with a sensitivity and specificity of 100% and 100% respectively.

Rapid Capture and Nondestructive Release of Extracellular Vesicles Using Aptamer-Based Magnetic Isolation.

Extracellular vesicles (EVs) play important roles in cell-cell communication by transferring cargo proteins and nucleic acids between cells. Due to their small size (50-150 nm) and low density, rapid capture and nondestructive release of EVs remains a technical challenge which significantly hinders study of their biofunction and biomedical application. To address this issue, we designed a DNA aptamer-based system that enabled rapid capture and nondestructive release of EVs in 90 min with similar isolation efficiency to ultracentrifugation (around 78%). Moreover, because we designed a DNA structure-switch process to release the exosomes, the isolated EVs maintained high bioactivity in cell-uptake assay and wound-healing assays. Using this method, we can isolate EVs from clinical samples and found that the amount of MUC1 positive EVs in breast cancer patient plasma sample is significantly higher than that in healthy donors. This DNA aptamer-based magnetic isolation strategy can be potentially applied for the biofunction study of EVs and EV-based point-of-care clinical tests.

Abstract P6-05-04: NEAT1 enhances drug sensitivity by inhibiting cancer stem-like cells in triple-negative breast cancer

Abstract Background: Accumulating evidence showed that long non-coding RNAs (lncRNAs) dysregulation is the hallmark of cancer. Nuclear paraspeckle assembly transcript 1 (NEAT1) has been reported to overexpress in many cancers, and promote cell growth and disease progression. However, the role of NEAT1 on drug sensitivity and stem-cell like property in triple-negative breast cancer is largely unknown. Methods: LncRNA expression profile were compared between breast cancer patients and healthy individuals using lncRNA array. Large scale validation of NEAT1 expression in blood samples were performed by real-time PCR. Triple-negative breast cancer (TNBC) cells, MDA-MB-231 and its cisplatin resistance subline (MDA-MB-231/cis) were used. Stable transfection of cells with NEAT1 knockdown by shRNA, and evaluated single cell clonogenic assay, aldehyde dehydrogenase (ALDH) activity, CD44+/CD24- and other cancer stem cell (CSC) markers. Drug sensitivity assay, flow cytometry analysis, immunofluorescence staining and xenograft model were used to assess the functional role of NEAT1. Results: Array data showed that NEAT1 was the top upregulated lncRNAs in the plasma of breast cancer patients. Consistent with the array data, validation of larger cohort of patients and healthy individuals (n=369) also demonstrated a higher expression of NEAT1 in breast cancer patients, in particular TNBC subtype. Knockdown of NEAT1 by shRNA sensitized breast cancer cells to cisplatin and taxol treatment. Cell proliferation and colony formation abilities were reduced with S-phase cell cycle arrest in shNEAT1 cells. Flow cytometry analysis revealed an induction of apoptosis with increased cleaved caspase-3. Cells expressing shNEAT1 abrogated ALDH activity, CD44+/CD24- subpopulation and expression of CSC markers (SOX2, NANOG and OCT4). More importantly, shNEAT1 cells retarded tumor growth in xenograft mice model and reduced CSC markers. Conclusion: Taken together, NEAT1 expression is differentially expressed in breast cancer patients, and particularly higher in patients with TNBC. These findings suggest a potent therapeutic target to improve drug sensitivity in patients with TNBC. Citation Format: Kwong A, Chen J, Siu J, Chuek I, Shin V. NEAT1 enhances drug sensitivity by inhibiting cancer stem-like cells in triple-negative breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P6-05-04.

Reversed phase UHPLC/ESI-MS determination of oxylipins in human plasma: a case study of female breast cancer.

The ultrahigh-performance liquid chromatography-mass spectrometry (UHPLC/MS) method was optimized and validated for the determination of oxylipins in human plasma using the targeted approach with selected reaction monitoring (SRM) in the negative-ion electrospray ionization (ESI) mode. Reversed phase UHPLC separation on an octadecylsilica column enabled the analysis of 63 oxylipins including numerous isomeric species within 12-min run time. The method was validated (calibration curve, linearity, limit of detection, limit of quantification, carry-over, precision, accuracy, recovery rate, and matrix effect) and applied to 40 human female plasma samples from breast cancer patients and age-matched healthy volunteers (control). Thirty-six oxylipins were detected in human plasma with concentrations above the limit of detection, and 21 of them were quantified with concentrations above the limit of quantitation. The concentrations determined in healthy controls are in a good agreement with previously reported data on human plasma. Quantitative data were statistically evaluated by multivariate data analysis (MDA) methods including principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA). S-plot and box plots showed that 13-HODE, 9-HODE, 13-HOTrE, 9-HOTrE, and 12-HHTrE were the most upregulated oxylipin species in plasma of breast cancer patients.

Study of the Differential Expression of Non-coding RNA, such as PRNCR1, in the Plasma of Breast Cancer Patients

Study of the Differential Expression of Non-coding RNA, such as PRNCR1, in the Plasma of Breast Cancer Patients

A Role for Tryptophan-2,3-dioxygenase in CD8 T-cell Suppression and Evidence of Tryptophan Catabolism in Breast Cancer Patient Plasma.

Tryptophan catabolism is an attractive target for reducing tumor progression and improving antitumor immunity in multiple cancers. Tumor infiltration by CD8 T cells correlates with improved prognosis in triple-negative breast cancer (TNBC) and a significant effort is underway to improve CD8 T-cell antitumor activity. In this study, primary human immune cells were isolated from the peripheral blood of patients and used to demonstrate that the tryptophan catabolite kynurenine induces CD8 T-cell death. Furthermore, it is demonstrated that anchorage-independent TNBC utilizes the tryptophan-catabolizing enzyme tryptophan 2,3-dioxygenase (TDO) to inhibit CD8 T-cell viability. Publicly available data revealed that high TDO2, the gene encoding TDO, correlates with poor breast cancer clinical outcomes, including overall survival and distant metastasis-free survival, while expression of the gene encoding the more commonly studied tryptophan-catabolizing enzyme, IDO1 did not. Metabolomic analysis, using quantitative mass spectrometry, of tryptophan and its catabolites, including kynurenine, in the plasma from presurgical breast cancer patients (n = 77) and 40 cancer-free donors (n = 40) indicated a strong correlation between substrate and catabolite in both groups. Interestingly, both tryptophan and kynurenine were lower in the plasma from patients with breast cancer compared with controls, particularly in women with estrogen receptor (ER)-negative and stage III and IV breast cancer. IMPLICATIONS: This study underscores the importance of tryptophan catabolism, particularly in aggressive disease, and suggests that future pharmacologic efforts should focus on developing drugs that target both TDO and IDO1.

Evaluation of miRNA-9 and miRNA-34a as potential biomarkers for diagnosis of breast cancer in Iranian women

PurposeMicroRNAs are involved in diverse biological processes and their dysregulation is a common event in various diseases including breast cancer. Breast cancer is a major threat to women's health. This study was designed to examine the expression levels of miR-9 and miR-34a in breast tumor tissue samples and plasma of breast cancer patients, compare their expression pattern between tissue samples and plasma samples of patients and analyze their relationship with tumor clinical features. Also, the potential of these miRNAs as diagnostic biomarkers for breast cancer was investigated. Materials and methodsThe expression levels of miR-9, miR-34a and CDH1 were measured by real-time reverse transcription polymerase chain reaction and ΔΔct method. Data were analyzed using t-test and one-way ANOVA. The sensitivity and specificity of miRNAs were determined by receiver operating characteristic (ROC) curve. Results and discussionThe expression levels of miR-9 and miR-34a were significantly down-regulated in tumor tissues compared to healthy tissues (fold change = 0.26, p = 0.0051 for miR-9 and fold change = 0.55, p = 0.021 for miR-34a). While no significant difference was observed in the expression levels of miR-9 (p = 0.205) and miR-34a (p = 0.132) in plasma samples of patients compared to normal plasma. CDH1 expression in tumor tissue was not significantly different from normal tissue (p = 0.33). We found that expression level of miR-9 in patients with tumor size larger than 5 cm (p = 0.026) and expression level of miR-34a in patients with higher stage (lll & lV, p = 0.03) were significantly down-regulated. Also miR-34a expression level was positively correlated with patient's age (p = 0.03). ConclusionAccording to the ROC curves, the area under the curve (AUC) of miR-9 in tissue was 0.71 (p = 0.009) with sensitivity 83.33% and specificity 70.37%. The AUC for miR-34a in tissue was 0.72 (p = 0.007) with sensitivity 72% and specificity 76%. Thus miR-9 and miR-34a have the capability for distinguishing tumor tissues from healthy tissues and the study of their expression levels in tissue may be used as a biomarker for the diagnosis of breast cancer patients from healthy women.

The expression pattern of miR-16 in plasma of breast cancer patients attending radiotherapy clinic in luth

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Abstract B63: Prognostic and predictive value of EGFR and EGFR-ligands in blood of breast cancer patients: A systematic review

Abstract The epidermal growth factor receptor (EGFR) and its ligands are involved in breast cancer pathogenesis and serve as co-targets for dual- and pan-EGFR-inhibitors in breast cancer treatment. There are indications that EGFR and EGFR-ligands are involved in resistance towards certain breast cancer treatments, including trastuzumab and antiendocrine treatment. The aim of the systematic review is to explore the validity of EGFR and EGFR-ligands in blood as prognostic and predictive biomarkers in breast cancer. The systematic review is conducted in accordance with the Preferred Reporting Items for Systematic Review and Meta-analysis Protocols (PRISMA-P) 2015 Statement. Literature searches were conducted in PubMed, Embase, and Cochrane Library to identify publications exploring correlations between levels of EGFR or EGFR-ligands in serum or plasma of breast cancer patients and prognostic or predictive outcome measures. A total of 2152 publications were identified and assessed for eligibility through title screening, followed by abstract screening and eventually assessment of full-text reports. A total of 16 publications were eligible for inclusion, of which 12 studies evaluated EGFR in serum and 5 studies evaluated one or more of the EGFR-ligands in serum. Current evidence indicates associations between low baseline serum-EGFR and shorter survival or reduced response to treatment in patients with advanced or metastatic breast cancer, especially in patients with estrogen and/or progesterone receptor-positive tumors. Though several studies did not find the same significant trends, none of the identified studies reported opposing results. Regarding EGFR-ligands, the prognostic and predictive value of S-Amphiregulin (S-AREG), S-Epidermal Growth Factor (S-EGF), and S-Transforming Growth Factor-α (S-TGFα) in breast cancer has been investigated in several studies, whereas no studies evaluated S-Betacellulin, S-Heparin Binding Epidermal Growth factor, S-Epiregulin, or S-Epigen. Most of the studies evaluating S-AREG, S-EGF, or S-TGFα included few patients and all studies were performed on patients experiencing HER2-positive advanced breast cancer (in one study HER2-status was not reported). Only one larger study was identified. In this study no associations between S-AREG, S-EGF, or S-TGFα and progression-free survival were observed in patients receiving either pertuzumab+trastuzumab+docetaxel or placebo+trastuzumab+docetaxel. However, decreases or increases in tumor marker levels were not evaluated in this study. The utility of both S-EGFR and serum levels of EGFR-ligands as predictive and prognostic markers in breast cancer has only been investigated in highly selected subsets of breast cancer patients, so extensive aspects remain to be uncovered. Based on the current knowledge, it is not possible to make recommendations either for or against clinical application. Large, well-planned studies are needed in order to clarify and validate the clinical impact of EGFR and EGFR-ligands in the clinical management of breast cancer patients. Citation Format: Ina Mathilde Kjaer, Troels Bechmann, Ivan Brandslund, Jonna Skov Madsen. Prognostic and predictive value of EGFR and EGFR-ligands in blood of breast cancer patients: A systematic review [abstract]. In: Proceedings of the AACR Special Conference: Advances in Breast Cancer Research; 2017 Oct 7-10; Hollywood, CA. Philadelphia (PA): AACR; Mol Cancer Res 2018;16(8_Suppl):Abstract nr B63.

Abstract LB-226: Exosomal liquid biopsy reveals mRNA and lincRNA biomarkers in early stage breast cancer patient plasma

Abstract Introduction Breast cancer is the most prevalent cancer in women, with approximately 250,000 diagnoses per year in the US. Non-invasive detection of breast cancer is of critical importance but has proven challenging due to the rate of false-positive diagnoses with current tests. Liquid biopsies including circulating tumor cells (CTCs) or cell-free DNA (cfDNA) have struggled with the detection of early stage disease. For example, a recent multi-analyte (cfDNA+protein) analysis, ‘cancerSEEK', achieved a sensitivity of just 33% in breast cancer, highlighting the challenges facing the development of more sensitive and specific diagnostics for this disease. Exosome-based liquid biopsy is a promising approach for minimally-invasive and highly sensitive diagnostics and it has been demonstrated that combining exosomal RNA and cfDNA greatly enhances mutation detection compared to profiling cfDNA alone. While most liquid biopsies profile mutations, studying RNA abundance in exosomes adds a new dimension to these non-invasive diagnostics. To date, much of the focus on exosomal RNA expression profiling has been on the small-RNA fraction. Here we demonstrate that whole-transcriptome profiling of mRNAs and lincRNAs greatly expands the landscape of potential biomarkers to clinically actionable genes. Results We have developed a novel platform designed to perform long-RNA sequencing on transcripts obtained from exosomes. We used this platform to compare expression profiles of total plasma exosomes versus subpopulations enriched for breast cancer-derived exosomes (CDE) versus depleted of non-cancer exosomes (NCE). The NCE-depleted and CDE-enriched exosomes equally outperformed total plasma exosomes, each detecting significantly more genes exhibiting breast cancer vs. healthy expression differences. We performed NCE-depletion on 1.5 mL of input plasma from 15 stage I & II ER+/Her2- breast cancer patients and 12 healthy women matched for age & menopausal-status. RNA-seq data from these samples detected over 10,000 mRNAs and over 1,000 lincRNAs. Of these, we observed significantly increased abundance in over 100 mRNAs and lincRNAs in these early stage breast cancer patients. These mRNAs are enriched for gene-sets including those previously implicated in ‘breast cancer', ‘chromatin remodeling', and ‘immune response'. We also performed RNA-seq on formalin-fixed paraffin-embedded (FFPE) samples from healthy and matched breast cancer tissue. The >100 genes found to be more abundant in breast cancer plasma exosomes significantly (p<0.05) separate the FFPE samples into two clusters corresponding to breast cancer patients and normal individuals, lending confidence to the exosomal signature. Conclusions This preliminary analysis highlights the exciting potential of exosomal long RNA based liquid biopsy for non-invasive early-stage detection of breast cancer. The platform is readily applicable to other disease areas and other biofluids such as urine or CSF. Citation Format: Sudipto K. Chakrabortty, Robert R. Kitchen, Christine M. Coticchia, Vasisht R. Tadigotla, Erez Eitan, Elena Castellanos-Rizaldos, Lisa Bedford, Sunita Badola, Michael D. Valentino, Nicholas Colafemina, Hidefumi Uchiyama, Mario Morken, Miguel Williams, Sylvie Vincent, Hadi Danaee, Seth Yu, Johan Skog. Exosomal liquid biopsy reveals mRNA and lincRNA biomarkers in early stage breast cancer patient plasma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-226.

Methylation of NBPF1 as a novel marker for the detection of plasma cell-free DNA of breast cancer patients

BackgroundRecent studies revealed that tumor-specific gene methylation can be detected in the circulating cell-free DNA (cfDNA) of cancer patients; therefore, methylated cfDNA is considered a promising biomarker. Human neuroblastoma breakpoint family member 1 (NBPF1) was originally identified in a neuroblastoma (NB) patient. The present study is the first to evaluate the presence of NBPF1 gene methylation in cfDNA in plasma of breast cancer patients. MethodsDifferentially methylated cfDNA was screened using bisulfite sequencing with a next-generation sequencer (BS-seq) among 25 breast cancer patients, 25 patients with a benign breast disease and 25 healthy female volunteers. Then, five specific methylation sites in the NBPF1 gene were verified in 11 breast cancer samples, and two further sites in the NBPF1 promoter were detected in 52 breast cancer patients (stages I-III), 31 patients with benign breast disease and 30 healthy controls by using methylation-specific PCR (MSP). Furthermore, the association between the methylation statuses of NBPF1 and the clinicopathological characteristics of breast cancer patients was analyzed. ResultsBS-seq demonstrated that the NBPF1 methylation levels in breast cancer patients were higher than those in patients with benign breast disease and healthy controls. The MSP results showed that the methylation rates of two sites in the NBPF1 promoter were 67.1% and 61.4% in breast cancer patients, 48.2% and 59.6% in patients with benign breast disease, and 40.9% and 48.1% in healthy controls, respectively. The methylation rates of one site were significantly different among the three groups (p < .05), with the highest rate in breast cancer patients. Moreover, there was no statistically significant correlation between the NBPF1 promoter methylation and the major clinicopathological features of the patients. ConclusionsThese results indicate that hypermethylation of the NBPF1 promoter occurs in a significant proportion of breast tumors and that NBPF1-methylated cfDNA may serve as a potential tumor marker for breast cancer.

Detection of marker miRNAs in plasma using SOI-NW biosensor

This work demonstrates the possibility of detection of DNA-oligonucleotide (oDNA) and microRNAs (miRNAs) extracted from blood plasma and associated with breast cancer (BC) in a buffer solution using the nanowire (NW) biosensor based on the silicon-on-insulator (SOI) structures. For biospecific detection, the NW surface was modified with oligonucleotide probes (oDNA-probes) complementary to the well-known miRNA sequence. It has been shown that the SOI-NW biosensor with immobilized oDNA-probes of this type can be used to detect complementary oDNA in a buffer solution with a concentration sensitivity of 10−17 М. The study also demonstrates the use of such biosensor for detection of an increased level of miRNAs isolated from the plasma of breast cancer patients. Application of the NW biosensor enables to distinguish between patients with a diagnosis of breast cancer from those with ovarian cancer (OC) by the value of an increased miRNAs level in the blood plasma of breast cancer patients.

Robust expression of tumor suppressor miRNA\u2019s let-7 and miR-195 detected in plasma of Saudi female breast cancer patients

BackgroundFemale breast cancer is frequently diagnosed at a later stage and the leading cause of cancer deaths world-wide. Levels of cell-free circulating microRNAs (miRNAs) can potentially be used as biomarkers to measure disease progression in breast cancer patients in a non-invasive way and are therefore of high clinical value.MethodsUsing quantitative RT-PCR, circulating miRNAs were measured in blood samples collected from disease-free individuals (n = 34), triple-negative breast tumours (TNBC) (n = 36) and luminal tumours (n = 57). In addition to intergroup comparisons, plasma miRNA expression levels of all groups were analyzed against RNASeq data from cancerous breast tissue via The Cancer Genome Atlas (TCGA).ResultsA differential set of 18 miRNAs were identified in the plasma of breast cancer patients and 10 miRNAs were uniquely identified based on ROC analysis. The most striking findings revealed elevated tumor suppressor let-7 miRNA in luminal breast cancer patients, irrespective of subtype, and elevated miR-195 in plasma of TNBC breast cancer patients. In contrast, hsa-miR-195 and let-7 miRNAs were absent from cancerous TCGA tissue and strongly expressed in surrounding non-tumor tissue indicating that cancerous cells may selectively export tumor suppressor hsa-miR-195 and let-7 miRNAs in order to maintain oncogenesis.ConclusionsWhile studies have indicated that the restoration of let-7 and miR-195 may be a potential therapy for cancer, these results suggested that tumor cells may selectively export hsa-miR-195 and let-7 miRNAs thereby neutralizing their potential therapeutic effect. However, in order to facilitate earlier detection of breast cancer, blood based screening of hsa-miR-195 and let-7 may be beneficial in a female patient cohort.

Down-regulation of traditional oncomiRs in plasma of breast cancer patients.

Deregulated expression of microRNAs has the oncogenic or tumor suppressor function in cancer. Since miRNAs in plasma are highly stable, their quantification could contribute to more precise cancer diagnosis, prognosis and therapy prediction. We have quantified expression of seven oncomiRs, namely miR-17/92 cluster (miR-17, miR-18a, miR-19a and miR-20a), miR-21, miR-27a and miR-155, in plasma of 137 breast cancer (BC) patients. We detected down-regulation of six miRNAs in patients with invasive BC compared to controls; however, only miR-20a and miR-27a down-regulations were statistically significant. Comparing miRNA expression between early and advanced stages of BC, we observed statistically significant decrease of miR-17 and miR-19a. We identified down-regulation of miR-17 and miR-20a in patients with clinical parameters of advanced BC (lymph node metastasis, tumor grade 3, circulating tumor cells, higher Ki-67-related proliferation, hormone receptor negativity and HER2 amplification), when compared to controls. Moreover, decreased level of miR-17 was found from low to high grade. Therefore, miR-17 could represent an indicator of advanced BC. Down-regulated miR-27a expression levels were observed in all clinical categories regardless of tumor progression. Hence, miR-27a could be used as a potential diagnostic marker for BC. Our data indicates that any changes in miRNA expression levels in BC patients in comparison to controls could be highly useful for cancer-associated pathology discrimination. Moreover, dynamics of miRNA expression changes could be used for BC progression monitoring.

Effects of CYP2D6 and CYP3A5 polymorphisms on tamoxifen and its metabolites in Thai breast cancer patients.

PurposeThis study aimed to determine the effects of CYP2D6 and CYP3A5 polymorphisms on the levels of tamoxifen (TAM) and its metabolites in the plasma of breast cancer patients. The protocol was designed to test the associations between CYP2D6, CYP3A5 genotypes and phenotypes (extensive metabolizer [EM], intermediate metabolizer [IM] and poor metabolizer [PM]) and TAM, N-desmethyl tamoxifen (NDMT), endoxifen (END) and 4-hydroxytamoxifen (4OHT) concentrations.Patients and methodsOne hundred and thirty-four Thai breast cancer patients from the Thai Tamoxifen Project undergoing TAM treatment who met the inclusion/exclusion criteria were recruited. Plasma samples were assessed for the concentrations of TAM and its metabolites using high-performance liquid chromatography. The data are presented as actual values and metabolic ratios (MR). The hypotheses were tested using Kruskal–Wallis or Mann–Whitney U test, including the simple main effects analysis.ResultsThe patients had stage 0–IV breast cancer. The mean age and body mass index were 51.6±11.6 years and 24.0±4.3, respectively. Also, 53.0% of them were premenopausal, 10.4% were perimenopausal and 36.6% were postmenopausal, while 23.1% were CYP2D6-EM/CYP3A5-EM and 20.9% carried only CYP2D6 and CYP3A5 incomplete alleles. The median concentrations of TAM, NDMT, END and 4OHT were 374.7 (interquartile range [IQR] 230.2) ng/mL, 1,064.9 (IQR 599.6) ng/mL, 54.5 (IQR 52.5) ng/mL and 5.0 (IQR 3.1) ng/mL, respectively. MR (TAM-NDMT) and MR (NDMT-END) were statistically different (p=0.013 and p=0.014, respectively), while MR (4OHT-END) was not statistically different within the CYP2D6 phenotype (p=0.594). MR (TAM-4OHT) was not statistically different within the CYP2D6 phenotype (p=0.079), but it was potentially different from CYP3A5-PM (p=0.056). None of the MR was statistically different within the CYP3A5 phenotype.ConclusionCYP2D6 polymorphisms appear to affect END concentration through an NDMT subpathway and potentially affect 4OHT concentrations through a 4OHT subpathway in CYP3A5-PM group.