Angiogenesis Markers Quantification in Breast Cancer and Their Correlation with Clinicopathological Prognostic Variables

Tumoural angiogenesis is essential for the growth and spread of breast cancer cells. Therefore the aim of this study was to assess the diagnostic performance of angiogenesis markers in tumours and there reflecting levels in serum of breast cancer patients. Angiogenin, Ang2, fibroblast growth factor basic, intercellular adhesion molecule (ICAM)-1, keratinocyte growth factor (KGF), platelet-derived growth factor-BB, and VEGF-A were measured using a FASTQuant angiogenic growth factor multiplex protein assay. We observed that breast cancer tumours exhibited high levels of PDGF-BB, bFGF and VEGF, and extremely high levels of TIMP-1 and Ang-2, whereas in serum we found significantly higher levels of Ang-2, PDGF-BB, bFGF, ICAM-1 and VEGF in patients with breast cancer compared to the benign breast diseases patients. Moreover, some of these angiogenesis markers evaluated in tumour and serum of breast cancer patients exhibited association with standard clinical parameters, ER status as well as MVD of tumours. Angiogenesis markers play important roles in tumour growth, invasion and metastasis. Our results suggest that analysis of angiogenesis markers in tumour and serum of breast cancer patients using multiplex protein assay can improve diagnosis and prognosis in this diseases

Baseline blood immunological profiling differentiates between Her2– breast cancer molecular subtypes: implications for immunomediated mechanisms of treatment response

Oana Tudoran,1,2,* Oana Virtic,2,* Loredana Balacescu,1,2 Carmen Lisencu,3 Bogdan Fetica,4 Claudia Gherman,1 Ovidiu Balacescu,1 Ioana Berindan-Neagoe1,2,5 1Department of Functional Genomics and Experimental Pathology, The Oncology Institute “Prof Dr Ion Chiricuţă”, 2Research Center for Functional Genomics, Biomedicine and Translational Medicine, Iuliu Hatieganu University of Medicine and Pharmacy, 3Department of Radiotherapy I, 4Department of Pathology, The Oncology Institute “Prof Dr Ion Chiricuţă”, 5Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania *These authors contributed equally to this work Purpose: Breast cancer patients’ response to treatment is highly dependent on the primary tumor molecular features, with triple-negative breast tumors having the worst prognosis of all subtypes. According to the molecular features, tumors stimulate the microenvironment to induce distinct immune responses, baseline immune activation being associated with higher likelihood of pathologic response. In this study, we investigated the deconvolution of the immunological status of triple-negative tumors in comparison with luminal tumors and the association with patients’ clinicopathological characteristics.Patients and methods: Gene expression of 84 inflammatory molecules and their receptors were analyzed in 40 peripheral blood samples from patients with Her2- primary breast cancer tumors. We studied the association of triple-negative phenotype with age, clinical stage, tumor size, lymph nodes, and menopausal status.Results: We observed that more patients with estrogen (ER)/progesterone (PR)-negative tumors had grade III, while more patients with ER/PR-positive tumors had grade II tumors. Gene expression analysis revealed a panel of 14 genes to have differential expression between the two groups: several interleukins: IL13, IL16, IL17C and IL17F, IL1A, IL3; interleukin receptors: IL10RB, IL5RA; chemokines: CXCL13 and CCL26; and cytokines: CSF2, IFNA2, OSM, TNSF13.Conclusion: The expression levels of these genes have been previously shown to be associated with reduced immunological status; indeed, the triple-negative breast cancer patients presented with lower counts of lymphocytes and eosinophils than the ER/PR-positive ones. These results contribute to a better understanding of the possible role of antitumor immune responses in mediating the clinical outcome. Keywords: triple-negative breast cancer, gene expression, molecular mechanism, immunological status, clinical outcom

A PROPOSED CURATION PROTOCOL FOR DISCOVERY CANCER POTENTIAL BIOMARKER CANDIDATES

Omics technologies generate a large amount of data of various types, and their huge diversity in postgenomic era imposes the need to identify a useful functional flow connected to other resources already existed to enable more accurate discovery and selection of cancer potential biomarker candidates selection. On the other hand the biomarker discovery and validation (BMDV) studies presumed long-term and cost experiments. In order to reduce expenses due to exploration of the overwhelming research steps and to optimize the BMDV experimental design, we proposed a curation protocol applicable to the discovery of potential cancer biomarker candidates

Multi-angle in situ dynamic light scattering at a neutron spin echo spectrometer

A new sample environment, called Bio-Oven, has been built for the Neutron Spin Echo (NSE) SpectrometerJ-NSE Phoenix. It provides active temperature control and the possibility to perform Dynamic Light Scattering(DLS) measurements during the neutron measurement. DLS provides diffusion coefficients of the dissolvednanoparticles and thus one can monitor the aggregation state of the sample on a time scale of minutes duringthe spin echo measurement times on the order of days. This approach helps to validate the NSE data or toreplace the sample when its aggregation state influences the spin echo measurement results. The new Bio-Ovenis an in situ DLS setup based on optical fibers decoupling the free space optics around the sample cuvettein a lightproof casing from the laser sources and the detectors. It collects light from three scattering anglessimultaneously. Six different values of momentum transfer can be accessed by switching between two differentlaser colors. Test experiments were performed with silica nanoparticles with diameters ranging from 20 nm upto 300 nm. Their hydrodynamic radii were determined from DLS measurements and compared with the onesobtained by a commercial particle sizer. It was demonstrated that also the static light scattering signal can beprocessed and gives meaningful results. The protein sample apomyoglobin was used for a long-term test and ina first neutron measurement using the new Bio-Oven. The results prove that the aggregation state of the samplecan be followed using in situ DLS along with the neutron measurement

In Situ Dynamic Light Scattering Complementing Neutron Spin Echo Measurements on Protein Samples

Monitoring the state of the sample on the minute-time scale is crucial in case of sensitive soft matter or biological samples, given that neutron spin echo measurements take up to several days. Moreover, there is no method to interpret the normalized intermediate scattering function obtained by neutron spin echo measurements if relevant sample properties change during the measurement. Dynamic light scattering provides information on the diffusion constant of particles in solution (biological macromolecules like proteins, protein aggregates, polymer particles, etc.) with average hydrodynamic radii in a broad range from a few nanometers up to several microns. This information can be obtained within a few minutes and it offers a good overview of the current sample state. Details on the novel in situ dynamic light scattering set-up with one fixed scattering angle and first results obtained on a molten globule state of apo-myoglobin are presented