Serum Of Breast Cancer Patients Research Articles

Effect of anti-antibodies to estradiol and progesterone on the concentration of hormones in the blood serum of healthy women and breast cancer patients

The relationship between autoantibodies specific to estradiol (Es)/ progesterone (Pg) and the concentrations of these hormones in the blood serum of healthy women and breast cancer patients has been previously revealed. Purpose : to study the association between anti-idiotypic antibodies to Es and Pg and the concentrations of these hormones in postmenopausal healthy women and breast cancer patients. Material and Methods . The levels of Es and Pg in the blood serum of 206 healthy women and 480 breast cancer patients were studied using competitive enzyme immunoassay. The levels of autoantibodies to Es and Pg and appropriate antiidiotypic antibodies in the same serums were studied using non-competitive enzyme immunoassay. Results. The high personal ratio of hormones (Pg/Es >4.0) was associated with the low breast cancer risk (OR=0.3; р <0.0001), high individual ratio of specific antibodies (IgA-Pg1/IgA-Es1>1.0; р =0.004) and high individual ratio of anti-idiotypic antibodies (IgG-Pg2/IgG-Es2>1.0; p<0.001) in healthy women, but not in breast cancer patients. Conclusion. The association between anti-idiotypic antibodies of G class specific to Es/ Pg and hormonal status was revealed in healthy women but not in breast cancer patients. Further studies on the mechanisms of immunoregulation of carcinogenesis are needed.

Predicting the presence of breast cancer using circulating small RNAs, including those in the extracellular vesicles.

Emerging evidence indicates that small RNAs, including microRNAs (miRNAs) and their isoforms (isomiRs), and transfer RNA fragments (tRFs), are differently expressed in breast cancer (BC) and can be detected in blood circulation. Circulating small RNAs and small RNAs in extracellular vesicles (EVs) have emerged as ideal markers in small RNA‐based applications for cancer detection. In this study, we first undertook small RNA sequencing to assess the expression of circulating small RNAs in the serum of BC patients and cancer‐free individuals (controls). Expression of 3 small RNAs, namely isomiR of miR‐21‐5p (3′ addition C), miR‐23a‐3p and tRF‐Lys (TTT), was significantly higher in BC samples and was validated by small RNA sequencing in an independent cohort. Our constructed model using 3 small RNAs showed high diagnostic accuracy with an area under the receiver operating characteristic curve of 0.92 and discriminated early‐stage BCs at stage 0 from control. To test the possibility that these small RNAs are released from cancer cells, we next examined EVs from the serum of BC patients and controls. Two of the 3 candidate small RNAs were identified, and shown to be abundant in EVs of BC patients. Interestingly, these 2 small RNAs are also more abundantly detected in culture media of breast cancer cell lines (MCF‐7 and MDA‐MB‐231). The same tendency in selective elevation seen in total serum, serum EV, and EV derived from cell culture media could indicate the efficiency of this model using total serum of patients. These findings indicate that small RNAs serve as significant biomarkers for BC detection.

Serum-derived factors of breast cancer patients with brain metastases alter permeability of a human blood\u2013brain barrier model

BackgroundThe most threatening metastases in breast cancer are brain metastases, which correlate with a very poor overall survival, but also a limited quality of life. A key event for the metastatic progression of breast cancer into the brain is the migration of cancer cells across the blood–brain barrier (BBB).MethodsWe adapted and validated the CD34+ cells-derived human in vitro BBB model (brain-like endothelial cells, BLECs) to analyse the effects of patient serum on BBB properties. We collected serum samples from healthy donors, breast cancer patients with primary cancer, and breast cancer patients with, bone, visceral or cerebral metastases. We analysed cytokine levels in these sera utilizing immunoassays and correlated them with clinical data. We used paracellular permeability measurements, immunofluorescence staining, Western blot and mRNA analysis to examine the effects of patient sera on the properties of BBB in vitro.ResultsThe BLECs cultured together with brain pericytes in transwells developed a tight monolayer with a correct localization of claudin-5 at the tight junctions (TJ). Several BBB marker proteins such as the TJ proteins claudin-5 and occludin, the glucose transporter GLUT-1 or the efflux pumps PG-P and BCRP were upregulated in these cultures. This was accompanied by a reduced paracellular permeability for fluorescein (400 Da). We then used this model for the treatment with the patient sera. Only the sera of breast cancer patients with cerebral metastases had significantly increased levels of the cytokines fractalkine (CX3CL1) and BCA-1 (CXCL13). The increased levels of fractalkine were associated with the estrogen/progesterone receptor status of the tumour. The treatment of BLECs with these sera selectively increased the expression of CXCL13 and TJ protein occludin. In addition, the permeability of fluorescein was increased after serum treatment.ConclusionWe demonstrate that the CD34+ cell-derived human in vitro BBB model can be used as a tool to study the molecular mechanisms underlying cerebrovascular pathologies. We showed that serum from patients with cerebral metastases may affect the integrity of the BBB in vitro, associated with elevated concentrations of specific cytokines such as CX3CL1 and CXCL13.

Dysregulated EGFR pathway in serum in early-stage breast cancer patients: A case control study

The epidermal growth factor receptor (EGFR) and its ligands are involved in cancer pathogenesis and they might serve as circulating biomarkers. The current study aims to investigate if abnormal pre-treatment serum levels of EGFR and EGFR ligands are present in women with early-stage breast cancer and if up- or downregulation of EGFR and EGFR ligands occur in defined patient subgroups. Pre-treatment serum samples were obtained from 311 women with newly diagnosed early-stage breast cancer and from 419 healthy women and analysed for EGFR and the ligands: Epidermal growth factor (EGF), heparin-binding epidermal growth factor (HBEGF), betacellulin (BTC), amphiregulin (AREG), and transforming growth factor α (TGF-α). Previously, age-dependent 95% reference intervals for EGFR and the EGFR ligands have been established based on the healthy women population. S-EGFR, S-EGF, S-HBEGF, S-AREG, and S-TGFα were all significantly different in women with breast cancer compared to healthy women (p < 0.05). Elevated S-EGFR, according to the reference intervals, was present in 11.3% of breast cancer patients, whereas decreased S-EGF was found in 11.6%. Elevated S-EGFR was associated with estrogen receptor positivity of tumor (ER+) and a subgroup of ER + breast cancer patients showed markedly elevated S-EGFR (>120 ng/mL).

Meme Kanseri Olan Hastalarda Serum Homoarjinin ve İskemi Modifiye Albumin Düzeyleri

Objectives: Homoarginine and ischemia modified albumin are among the molecules that are intensely studied on. Homarginine and ischemia-modified albumin levels in breast cancer patients are extremely important in understanding the pathogenesis of diseases, associating these molecules with treatment, developing new treatment protocols, and even eliminating risk factors in healthy people before diseases occur. Our aim in this study is to determine homoarginin and ischemia-modified albumin levels in the serum of breast cancer patients and its relation with the disease. Material and Methods: 27 controls, 27 breast cancer were included in the study. Serum methylarginine and homoarginine levels were studied by mass spectrometric method, and ischemia modified albumin levels by colorimetric method in Selcuk University Faculty of Medicine Biochemistry Laboratory. Results: While the levels of L-monomethylarginine, Total Methyl Arginine, Homoarginine statistically decreased, asymmetric dimethylarginine and ischemia modified albumin levels significantly increased in patients compared to controls (p &lt;0.05). Conclusions: Increased ischemia modified albumin levels may indicate the oxidative stress that occurs in breast cancer. Increase in asymmetric dimethylarginine levels can lead to decreased nitric oxide synthesis and deterioration of the vascular structure. Keywords: Breast cancer; Homoarginine; Ischemia modified albumin; Mass spectrometry

Autophagy-associated circRNA circCDYL augments autophagy and promotes breast cancer progression

BackgroundAlthough both circular RNAs (circRNAs) and autophagy are associated with the function of breast cancer (BC), whether circRNAs regulate BC progression via autophagy remains unknown. In this study, we aim to explore the regulatory mechanisms and the clinical significance of autophagy-associated circRNAs in BC.MethodsAutophagy associated circRNAs were screened by circRNAs deep sequencing and validated by qRT-PCR in BC tissues with high- and low- autophagic level. The biological function of autophagy associated circRNAs were assessed by plate colony formation, cell viability, transwells, flow cytometry and orthotopic animal models. For mechanistic study, RNA immunoprecipitation, circRNAs pull-down, Dual luciferase report assay, Western Blot, Immunofluorescence and Immunohistochemical staining were performed.ResultsAn autophagy associated circRNA circCDYL was elevated by 3.2 folds in BC tissues as compared with the adjacent non-cancerous tissues, and circCDYL promoted autophagic level in BC cells via the miR-1275-ATG7/ULK1 axis; Moreover, circCDYL enhanced the malignant progression of BC cells in vitro and in vivo. Clinically, increased circCDYL in the tumor tissues and serum of BC patients was associated with higher tumor burden, shorter survival and poorer clinical response to therapy.ConclusionscircCDYL promotes BC progression via the miR-1275-ATG7/ULK1-autophagic axis and circCDYL could act as a potential prognostic and predictive molecule for breast cancer patients.

Abstract P3-01-03: Modulating inflammation with nucleic acid scavengers to prevent breast cancer metastasis

Abstract Rationale: Chronic inflammation is a hallmark of cancer. Inflammation, whether it is an underlying baseline condition or induced/exacerbated due to treatment, is a major hurdle that ultimately impacts the efficacy of standard of care therapy. However, no strategy exists to effectively control endogenous or therapy-induced inflammation in women with breast cancer. Moreover, there is limited information on how endogenous or therapy-induced inflammation modulates immune cells in women with breast cancer. Nucleic acid-containing pro-inflammatory complexes, termed Nucleic Acid-containing Damage Associated Molecular Patterns (NA-DAMPs), are released from dead or dying cancer cells or activated immune cells. NA-DAMPs activate nucleic acid sensing toll-like receptors (TLR) and other pattern recognition receptors (PRR). NA-DAMPs promote tumor progression and metastasis by inducing pro-tumorigenic signaling in cancer cells. Excessive and persistent NA-DAMP-mediated PRR signaling in immune cells is associated with immune dysfunction. Nucleic acid binding polymers have been used as vehicles for DNA/RNA delivery. We have discovered that a subset of nucleic acid binding polymers, termed nucleic acid scavengers (NAS), act as anti-inflammatory agents by capturing cancer-promoting pro-inflammatory NA-DAMPs. Therefore, we investigated if a NAS could limit cancer progression and metastasis by capturing and mitigating the effects of NA-DAMPs in immunocompetent murine models of breast cancer. Methods: We tested the use of the NAS PAMAM-G3 in the 1] orthotopic surgical resection 4T1 breast cancer model; 2] intravenous experimental metastasis 4T1 breast cancer model; 3] MMTV-PyMT spontaneous breast cancer model. We also measured changes in inflammation/inflammatory cell profile in tumor and in blood. Next, we investigated the relationship between therapy, NA-DAMPs and immune cell profile in blood from women with breast cancer. We also examined if NA-DAMPs in patient serum induces tumor invasion and if the NAS PAMAM-G3 prevents tumor invasion using an in vitro assay. Results: We have determined that the NAS PAMAM-G3 controls lung metastasis in immunocompetent murine models of breast cancer and our data indicate that: 1] chronic inflammation and inflammatory cells in the tumor and periphery promote cancer invasion and metastasis; 2] modulating inflammation by removing excessive NA-DAMPs using the NAS PAMAM-G3 limits cancer metastasis; 3] scavenging inflammation-associated NA-DAMPs significantly alters immune cell and cytokine/chemokine profile in the tumor and periphery, promotes T cell infiltration in the tumor and induces antitumor CD8+ T cell responses. In experiments conducted using blood collected longitudinally from women undergoing standard of care therapy, we observed that: 1] NA-DAMPs and inflammatory monocytes are elevated in women undergoing standard of care therapy; 2] Post-chemotherapy breast cancer patient serum increases invasion/migration of human breast cancer cells in vitro; 3] PAMAM-G3 abrogates increase in invasion/migration of human breast cancer cells in vitro in response to post-chemotherapy breast cancer patient serum. Conclusions: These results highlight the potential utility of NAS to treat cancer associated inflammation and thereby inhibit cancer metastasis by 1) limiting extracellular NA-DAMP-induced persistent activation of pro-inflammatory signaling, and 2) promoting full engagement of both the innate and adaptive immune systems. These principles will aid in the development of a novel anti-metastatic therapy for women with breast cancer. This work is supported by Department of Defense Breast Cancer Research Program Awards W81XWH-16-1-0512 (Nair) and W81XWH-16-1-0513 (Sullenger). Citation Format: Smita Nair, Eda Holl, Karenia Landa, Victoria Frazier, Elias Eteshola, David Boczkowski, Rachel Rempel, Shelley Hwang, Bruce Sullenger. Modulating inflammation with nucleic acid scavengers to prevent breast cancer metastasis [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P3-01-03.

Investigating the Potential and Pitfalls of EV-Encapsulated MicroRNAs as Circulating Biomarkers of Breast Cancer.

Extracellular vesicles (EVs) shuttle microRNA (miRNA) throughout the circulation and are believed to represent a fingerprint of the releasing cell. We isolated and characterized serum EVs of breast tumour-bearing animals, breast cancer (BC) patients, and healthy controls. EVs were characterized using transmission electron microscopy (TEM), protein quantification, western blotting, and nanoparticle tracking analysis (NTA). Absolute quantitative (AQ)-PCR was employed to analyse EV-miR-451a expression. Isolated EVs had the appropriate morphology and size. Patient sera contained significantly more EVs than did healthy controls. In tumour-bearing animals, a correlation between serum EV number and tumour burden was observed. There was no significant relationship between EV protein yield and EV quantity determined by NTA, highlighting the requirement for direct quantification. Using AQ-PCR to relate miRNA copy number to EV yield, a significant increase in miRNA-451a copies/EV was detected in BC patient sera, suggesting potential as a novel biomarker of breast cancer.

Proteomic Analysis of Inflammatory Biomarkers Associated With Breast Cancer Recurrence.

Breast cancer is the most frequent cancer detected for women, and while our ability to treat breast cancer has improved substantially over the years, recurrence remains a major obstacle. Standard screening for new and recurrent breast cancer involves clinical breast imaging. However, there is no clinically approved noninvasive body fluid test for the early detection of recurrent breast cancer. Materials and Method: In this study, we analyzed serum samples from both recurrent and nonrecurrent breast cancer patients by different proteomics methods to identify biomarkers in patients with recurrence of disease. Comparative data analysis identified several histone deacetylase (HDAC) proteins, which were found at significantly higher levels in the serum of recurrent breast cancer patients: HDAC9 (C-term) (P=0.0035), HDAC5 (C-term) (P=0.013), small ubiquitin-like modifier 1 (N-term) (P=0.017), embryonic stem cell-expressed Ras (inter) (P=0.018), and HDAC7 (C-term) (P=0.020). Chronic inflammation plays a critical role in the development of the breast cancer recurrence, and we identified several proinflammatory cytokines that were present at elevated levels only in recurrent breast cancer patient serum. Our data indicated that the epigenetic regulation of inflammatory processes plays a critical role in breast cancer recurrence. The identified proteins could lay the groundwork for the development of a serum-based breast cancer recurrence assay.

Estimation of trace elements (selenium, iron) and their biological effect in serum levels of breast cancer patients

Background: Breast cancer (BC) is the most commonly occurring cancer that records a significant percentage of all cancers among women. The most common type of BC is ductal carcinoma. The trace elements, selenium (Se) and iron (Fe), have an important role in the metabolism of human body in which they have very vital biological effect on the enzymes involved in antioxidants and immune system. Materials and Methods: Venous blood samples were collected into plain vials from 50 patients and 50 controls, which were transported to the laboratory on ice and centrifuged at 3000 rpm for 15 min, and the serum was stored at −4°C until the day of mineral estimation, using SHIMADZU Atomic Absorption Spectroscopy AA7000 using Graphite Furnace Technique. Results: There was a significant (P 0.05) in the serum of BC patients as compared with normal healthy controls. Conclusion: We found a significant association between trace elements (serum Se and Fe) with BC.

The Diagnostic Value of Serum miR-129 in Breast Cancer Patients with Bone Metastasis

The current study aims to investigate the expression of serum miR-129 in breast cancer patients with bone metastasis and to further study its diagnostic role. Serum samples of 60 patients with bone metastasis of breast cancer and 60 patients with non-bone metastasis of breast cancer were collected. The expression of serum miR-129 in breast cancer patients was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). The relationship between miR-129 and bone metastasis or bone pain in breast cancer patients with bone metastasis was analyzed. Receiver operating characteristic (ROC) curve was performed to analyze the diagnostic role of serum miR-129 expression in breast cancer patients with bone metastasis. The expression of miR-129 in serum of breast cancer patients with bone metastasis was significantly lower than that of non-bone metastasis patients. Furthermore, the lower the expression level of serum miR-129 was, the higher the degree of bone metastasis and bone pain was found in breast cancer patients. The ROC curve showed that the area under the curve (AUC) of miR-129 expression in serum for diagnosis of bone metastasis of breast cancer was 0.941 (95% CI: 0.891 - 0.991) with the sensitivity and specificity of 87.5% and 91.7%, respectively. In summary, decreased serum miR-129 in breast cancer patients can be used as a potential diagnostic marker of bone metastasis in breast cancer patients.

Detection of breast cancer based on novel porous silicon Bragg reflector surface-enhanced Raman spectroscopy-active structure

In this Letter, the surface-enhanced Raman scattering (SERS) signal of early breast cancer (BRC) patient serum is obtained by a composite silver nanoparticles (Ag NPs) PSi Bragg reflector SERS substrate. Based on these advantages, the serum SERS signals of 30 normal people and 30 early BRC patients were detected by this substrate. After a baseline correction of the experimental data, principal component analysis and linear discriminant analysis were used to complete the data processing. The results showed that the diagnostic accuracy, specificity, and sensitivity of the composite Ag NPs PSi Bragg reflector SERS substrate were 95%, 96.7%, and 93.3%, respectively. The results of this exploratory study prove that the detection of early BRC serum based on a composite Ag NPs PSi Bragg reflector SERS substrate is with a stable strong SERS signal, and an unmarked and noninvasive BRC diagnosis technology. In the future, this technology can serve as a noninvasive clinical tool to detect cancer diseases and have a considerable impact on clinical medical detection.

Isolation of Small Extracellular Vesicles From Human Serum Using a Combination of Ultracentrifugation With Polymer-Based Precipitation.

Methods for reproducibly isolating and enriching small extracellular vesicles (EVs) from blood are essential for clinical utilization of small EVs in cancer patients. We combined ultracentrifugation (UC) with polymer-based precipitation (ExoQuick [EQ] or Total Exosome Isolation [TEI] kit) to isolate small EVs (diameter, 30–150 nm) from the serum of breast cancer patients. We compared the performance of four cycles of UC (UC4x) with that of two cycles of UC followed by enrichment using the EQ (UC2x→EQ) or TEI (UC2x→TEI) kits. The mean concentration of small EVs isolated from 1 mL of serum using UC2x→EQ (139.0±29.1 µg) and UC2x→TEI (140.4±5.0 µg) did not differ from that obtained using UC4x (141.8±26.9 µg). The mean number of EV particles obtained using UC4x was 29.2±9.9×109 per mL of serum, whereas UC2x→EQ and UC2x→TEI yielded higher numbers of EVs (50.7±17.0×109 and 59.3±20.6×109, respectively). Concentrations of EV microRNAs, including miR-21 and miR-155, did not differ between the three methods. In conclusion, performing UC prior to the use of polymer-based precipitation kits could be feasible for isolating small EVs from human serum in large sample-based translational researches.

PIXE analysis of blood serum of breast cancer patients undergoing successive chemotherapy

Particle induced X-ray emission technique was used to obtain the serum elemental profile of healthy subjects and breast cancer patients (BCPs) undergoing chemotherapy. Concentrations of elements K, Ca, Ti, V, Cr, Mn, Fe, Co, Cu, Zn, Se, Br, Rb, and Sr were determined. The increased levels of Ca, Cr, Fe and Cu and diminished levels of Ti, Zn, and Se observed in BCPs prior to therapy, were restored to near normal levels after the fifth cycle of chemotherapy. This work elicits a unique possibility of identifying novel trace element based anti-tumour drugs for breast cancer.

Evaluation of microRNA-9 and -192 expression levels as biomarkers in patients suffering from breast cancer.

Given the global outbreak of breast cancer and its debilitating effect on women's health, it is not surprising that tremendous efforts have been made with an aim of shedding more light on the mechanisms involved in the pathogenesis of this type of cancer. Among the long list of risk factors associated with this malignancy, recently, the role of microRNAs (miRNAs or miRs) has turned into a hotspot for breast cancer investigations. miRNAs approximately 20 nucleotides in length and are located in either an exon or an intron, playing a role in the regulation of gene expression. In the present study, we extracted RNA from both the serum and cancerous tissue of breast cancer patients and after synthesizing the cDNA, we performed quantitative PCR to determine the expression levels of miR-9 and miR-192. The resulting data revealed that while the mRNA expression level of miR-9 was significantly decreased in the breast cancer tissues, there was no noticeable change in the expression level of this miRNA in the serum samples. Likewise, we found that the marked downregulation of miR-192 was only restricted to the cancerous tissues, but was not found in the serum of patients. Based on the meaningful downregulation of the expression of miR-9 and miR-192, this study provides a plausible framework for these miRNAs as effective biomarkers for breast cancer patients.

Reflecting Size Differences of Exosomes by Using the Combination of Membrane-Targeting Viscosity Probe and Fluorescence Lifetime Imaging Microscopy.

Exosomes are cell-secreted membrane-coated vesicles with their sizes variable from 30 to 150 nm. So far, there is no simple, fast, and economical way to evaluate the sizes of exosomes in living systems. Here, we put forward a hypothesis in which the sphere sizes (resulting in different curvature) may affect the local mobility/viscosity of exosome membranes. Based on this hypothesis, we propose a novel method to evaluate the exosome sizes by quantifying the membrane viscosity. For this sake, we design a membrane-targeting molecular rotor with its fluorescence lifetime sensitive to viscosity and use it under a fluorescence lifetime imaging microscope (FLIM). Through a multiple-step ultrafiltration technique, we isolate three individual size distributions (10-50, 50-100, and 100-220 nm) with exosomes from HeLa and MCF-7 cell culture media and then perform the FLIM assay on the above two groups. In both cases, we indeed find a regular pattern in which the membrane viscosity reflected by lifetime decreases with exosome sizes. We then perform the assay on exosomes from cancer cells, corresponding normal tissue cells, and serum of breast cancer patients. We find that exosomes from cancer cells have a fluorescence lifetime (larger viscosity) longer than that of normal tissue cells. The average fluorescence lifetime of exosomes from a triple-negative breast cancer patient is longer (or the viscosity is larger) than that of a HER2 positive one. Therefore, our new and simple method may hold application prospects in future cancer diagnosis.

The polymorphism of Apelin gene rs2235306 and serum Apelin level in breast cancer women in Samarra- Iraqi population

Breast cancer (BC) is the most common cancer leading to death in women and the fifth type of cancer leading to death worldwide, The study was done on 59 women, including 20 non-breast cancer women as a control group, their ages ranged from (30-74years) and 39 women with breast cancer their ages ranged from (29-80years).The results showed significant increase (p ≤0.05) in the concentration of Apelin in women with BC compared with non-breast cancer women, This finding suggests that Apelin may promoted carcinogenesis. The result of Apelin rs2235306 polymorphism using T-ARMS-PCR technique show no difference in the allele frequencies between BC patients and control group, the results also show that there is no association of the observed genotypes (TT&amp;TC) with serum Apelin concentration in the studied Population. The result of the study suggest that the serum Apelin level can be independent of Apelin rs2235306 polymorphism which should be confirmed by further studies on larger populations.&#x0D; The aim: The aim of this study was to determine Apelin level in serum and apln rs2235306 polymorphism in breast cancer (BC) patients in Samarra city-Iraq.

Based on magnetic beads to develop the kit for extraction of high-quality cell-free DNA from blood of breast cancer patients

Early detection of breast cancer (BC) is very important to avoid mortality caused by it. It is concluded in recent studies that the DNA in patient's serum can be used as biomarker for cancer detection. Therefore, extracting high-quality DNA from patient's serum can achieve this goal. In this work, we compared three methods (phenol-chloroform, column-based and magnetic beads) of extracting DNA from serum by means of purchased commercial kits and concluded that column-based kit is the best one to extract Cell-free DNA (cfDNA). Then we developed a new kit named PuriMag magnetic beads kit to extract DNA from serum of BC patients. Further, we described the properties of this new kit including its recovery capacity and stability. We compared the extraction performances of PuriMag magnetic beads kit and column-based kit and concluded that PuriMag magnetic beads are a better method to extract cfDNA. Surely PuriMag magnetic beads also suit automation and will be a useful method for fast, high-quality and automatic DNA extraction from serum.

High Levels of Cartilage Oligomeric Matrix Protein in the Serum of Breast Cancer Patients Can Serve as an Independent Prognostic Marker.

Background: Cartilage oligomeric matrix protein (COMP) is a pentameric cartilage protein also expressed in breast cancer tumors. A high expression of COMP evaluated by immunohistochemical staining is as an independent prognostic marker associated with poor patients' prognosis.Methods: Herein, levels of COMP were analyzed using an IVD approved ELISA in serum samples from 233 well-characterized breast cancer patients; 176 with metastatic breast cancer; and 57 in an early stage of the disease.Results: The metastatic patients had double the concentration of serum COMP compared with those with early breast cancer. High levels of COMP in sera of metastatic patients were associated with the histological subtype (p = 0.025) and estrogen receptor positivity (p = 0.019) at the time of breast cancer diagnosis. Further, correlation was observed between the serum levels of COMP and the presence of liver (p = 0.010) or bone (p = 0.010) metastases in this population. Most importantly, elevated serum levels of COMP appear to serve as an independent prognostic marker of survival as assessed by Cox proportional hazard regression analysis (p = 0.001) for the metastatic patients. Among metastatic patients treated with taxanes (Docetaxel-Paclitaxel) as part of their first metastatic line (n = 25), those with high levels of serum COMP detected in the metastatic stage of the disease had a shorter median survival (0.2 years) compared with those with low levels of serum COMP (1.1 years) (p = 0.001).Conclusions: Taken together, the serum levels of COMP are elevated in the metastatic patients and may be a potential novel biomarker for the evaluation of the prognosis in this population.

269P - Predicting the presence of breast cancer using circulating small RNA in the serum

BackgroundSmall RNAs are multiple classes of short non-coding RNAs (ncRNAs) that include microRNAs (miRNAs), transfer RNA fragments (tRFs), and other ncRNA fragments that play essential role in gene regulation. Small RNAs exist in blood circulation within extracellular vesicles (EVs), which have been the center of the attention at cell-to-cell communication tools, or bound to proteins or lipid. Recent studies have shown that the expression levels of small RNAs are different between patients with breast cancer (BC) and cancer-free individuals. Thus, circulating small RNAs have attracted as valuable biomarkers for cancer detection. MethodsFirst, we assessed and the expression level of circulating small RNAs in the serum of BC patients (n=78) and cancer-free volunteers (control) (n=72) using next generation sequencer. By comparing those expression level, we identified some small RNAs that have signifficant difference between 2 groups as biomarker candidates for BC detection. We constructed a diagnostic model using some small RNAs from biomarker candidates. To test the possibilities that those candidates are released from cancer cells, we next profiled small RNAs within EVs that isolated from the serum of participants in this study and from breast cancer cells (MCF-7 and MDA-MB-231) and normal epithelial cells (184-h TERT). ResultsTwelve circulating small RNAs that expressed significantly higher in BC patients compared to control were identified, indicating potential biomarkers for BC detection. A diagnostic model using 4 small RNAs including isoforms of miRNAs (isomiRs) and tRFs was. The ROC curve analysis revealed that our model showed a high diagnostic accuracy of AUC 0.945 and achieved discriminating stage 0 BCs from control. Importantly, significantly different expression in serum-EVs was observed in 4 small RNAs between BC patients and control. Furthermore, 4 small RNAs were also observed in EVs derived from cell culture media in breast cancer cells and normal epithelial cells and those expressions were aberrant as the same as those of serum-EVs. ConclusionsThese findings suggests circulating small RNAs in serum serve as potential biomarkers for BC detection that enables to include the molecular movement of cancer cells. Legal entity responsible for the studyHidetoshi Tahara. FundingHas not received any funding. DisclosureH. Tahara: Advisory / Consultancy: MiRTeL.