BackgroundMinimizing the effects of transportation on the properties of biological material is a major challenge for the scientific community. The viability of cells is important in cases where their study is urgent for evaluation of treatment response or for the study of cancer progression. Circulating tumor cells (CTCs) constitute a cell subpopulation with great importance for oncologists, because of their prognostic value. Detection and isolation of CTCs from blood samples is a routine activity in many laboratories, but concerns exist with regard to the maintenance of the cells during transportation. In this study, experiments were conducted to determine the stability of gene and protein expression in CTCs over a period of 96 h.ResultsBlood samples collected from healthy individuals and patients with cancer were each divided into five aliquots, which were stored at 2–8 °C and analyzed after 0, 24, 48, 72 and 96 h of storage. CTCs from patients and CD45-negative cells from healthy individuals were isolated each day using enrichment protocols, and qPCR was performed to determine expression levels of genes encoding specific biological markers. In addition, cells from breast and colon cancer cell lines were spiked into blood samples from healthy individuals, and these samples were stored and analyzed over a period of 96 h by PCR and by flow cytometry. The markers that were studied included housekeeping genes and genes associated with the response to chemotherapy, as well as genes encoding transcription factors. The results demonstrated that the expression profiles of specific genes and proteins in CTCs were not significantly affected by 72 h of storage. After 96 h of storage, expression of some genes was altered.ConclusionThe transportation of blood at low temperature (2–8 °C) in the presence of the anticoagulant EDTA can protect CTCs from alteration of gene and protein expression for at least 72 h. Furthermore, under these conditions, CTCs can be detected and isolated 96 h after blood collection.

Much of the specification for the basic embryonic body plan is the result of a hierarchy of developmental decisions at different developmental times. The extracellular matrix (ECM) appears to be a very dynamic structure during embryogenesis. One of the mesenchymal ECM proteins, tenascin, is reported to be transiently expressed during embryonic tissue development, and is absent or much reduced in most fully developed organs. The respiratory system is an outgrowth of the ventral wall of the foregut, and the epithelium of the larynx, trachea, bronchi and alveoli is of endodermal origin. The cartilaginous and muscular components are of mesodermal origin. The aim of this study was to investigate the role of tenascin-C (TNC) in the developing human lung, during the pseudoglandular, canalicular and saccular stage of lung maturation. Formalin-fixed, paraffin-embedded tissue from the lungs of 30 embryos (10 corresponding to the 10th to the 16th gestational week (pseudoglandular stage), 10 to the 17th to the 23rd gestational week (canalicular stage), and 10 to the 24th to the 27th gestational week (saccular stage), were investigated by conventional histology and immunohistology for the expression levels of TNC. The changes observed in the distribution patterns suggest that during embryogenesis, the rate of tenascin synthesis changes significantly. During the pseudoglandular stage, the density of cells expressing TNC was higher in the condensing mesenchyme surrounding the epithelial glands than in the epithelial cells, whereas the inverse result was observed during the canalicular stage. During the saccular stage the pattern of immunoreactivity with TNC was lower than those of the pseudoglandular and canalicular stage, either in epithelial or mesenchymal cells, but it was highly expressed in the basement membranes. This restricted spatiotemporal distribution suggests that tenascin has a key role (1) in mesenchymal tissue remodeling during the pseudoglandular stage, a period that describes the development of the complete bronchial tree and (2) on the epithelial cell shape and function during the canalicular stage, a period that describes the formation of pneumocytes type I and pneumocytes type II. The later, will produce the surfactant, a phospholipid-rich fluid capable of lowering surface tension at the air-alveolar interface. During the saccular stage, tenascin was present mainly in the basement membranes surrounding the acinar and vascular structures, indicating a supporting and mechanical role.

Circulating tumor cells are cells that detach from the primary tumor site and migrate to the bone marrow or other tissues where they can initiate a metastatic site. Liquid biopsies are an emerging tool in the past decades that enables us to detect Circulating Tumor Cells in patients' blood. Flow cytometry is a powerful tool used in liquid biopsy diagnostics. This aims to prove the sensitivity and specificity of a flow cytometric panel for the detection of CTCs in breast cancer patients using healthy individuals' samples as controls. The study was blinded to the data analyzing researcher. Statistical analysis followed and results show 86.9% area under the curve which indicates that the particular method can be very promising for diagnosing breast cancer.

Hormone-refractory prostate carcinoma has a different cell surface protein profile than hormone-sensitive prostate carcinoma, which provides migration ability and interactions with organs/tissues. Detection and association of these proteins with lymph node metastasis via lymphadenectomy might be beneficial for patients. Gene expression analysis in hormone-refractory and hormone-sensitive commercial cancer cell lines was performed and, after co-cultivation with osteoblasts or endothelial cells, knockdown experiments followed to validate potential biomarkers. “ Myeloid-associated differentiation markers, myosin 1b and phosphatidylinositol-4-phosphate-5-kinase type 1 alpha are implicated in metastasis”, their knockdown altered the expression of key regulators of endothelial-mesenchymal transition, invasion, motility and migration. In primary prostate tumors, these genes could be an indicator for future metastasis into lymph nodes.

e14656 Background: Lung cancer is the second most common type of cancer in both genders; however is the leading cause of cancer deaths. Adenocarcinoma constitutes more than 60% of non-small cell lung cancer (NSCLC), and accounts for approximately 40% of lung cancers. Therefore, early prediction, prognosis of the disease and treatment strategy is of primary importance. Molecular biology assays, including microarrays, enable identification of new targets. The above targets can be used either as biomarkers, useful for early prediction and prognosis of the disease, or as drugable targets, upon proof of their biological action. The present study aimed to identify new biomarkers and/or targets in lung cancer. Methods: Whole genome gene expression microarrays have been performed for 12 lung cancer cell lines (provided be ECACC, ATCC), representing non-small cell lung (adenocarcinoma, squamous, large cell) and small cell lung cancer. Real-time experiments were followed for validation of thirty genes that were overexpressed. The experiments were performed including other cell lines, representing breast, colon, prostate, hepatocellular, melanoma and pancreatic cancer. Sample from healthy donors, as well as normal lung epithelial cell were used. Results: Among the genes that were studied, there was observed downregulation or none expression in normal donors as well as in normal lung epithelia cells. VSIG10 overexpressed in adenocarcinoma and large cells, while no expression observed in breast, prostate, melanoma and pancreatic cancer. DCBLD2 overexpressed in all NSCLC samples, but not in small cells. LIFR was overexpressed mainly in squamous cells, while no expression or downregulation observed in the other types of cancer. The same expression profile was mentioned for PDGFC. Conclusions: Taking everything into consideration, it is obvious that the identification of new biomarkers and/or targets for lung cancer is of primary importance. The present study revealed different genes, with different biological function that might be used primary as biomarkers. Their role need to be clarified also in clinical samples as to be used as drugable targets. However, the above data are encouraging for identification of new therapeutic models in lung cancer.