Presence Of Cancer Cells Research Articles

Synchronized Rayleigh and Raman scattering for the characterization of single optically trapped extracellular vesicles

Extracellular Vesicles (EVs) can be used as biomarkers in diseases like cancer, as their lineage of origin and molecular composition depend on the presence of cancer cells. Recognition of tumor-derived EVs (tdEVs) from other particles and EVs in body fluids requires characterization of single EVs to exploit their biomarker potential. We present here a new method based on synchronized Rayleigh and Raman light scattering from a single laser beam, which optically traps single EVs. Rapidly measured sequences of the Rayleigh scattering amplitude show precisely when an individual EV is trapped and the synchronously acquired Raman spectrum labels every time interval with chemical information. Raman spectra of many single EVs can thus be acquired with great fidelity in an automated manner by blocking the laser beam at regular time intervals. This new method enables single EV characterization from fluids at the single particle level.

The Application of Levine’s Conservation Model on Nursing Care of Children with Cancer Experiencing Chemotherapy-Induced Mucositis in Indonesia

Background: Mucositis is a common side effect in children with cancer experienced chemotherapy. Levine’s Conservation Theory views children as open individuals who always respond to the environment. Children with cancer undergoing chemotherapy are seen as individuals who adapt to threats from the internal-external environment. The threat from the internal environment is the presence of cancer cells that threaten normal cells, while the external environment is the side effects of chemotherapy and environmental exposure. The nurse is responsible for carrying out a series of nursing processes to prevent the occurrence of mucositis due to chemotherapy.
 Objectives: Thisstudyaimedtoexplorethenursingprocessthatwasgiventochildrenexperiencing chemotherapy-induced mucositis using Levine’s Conservation Theory.
 Methods: This study was conducted with a case study. Ten children with cancer who experienced chemotherapy due to mucositis was participated in this study. Nursing process was applying by Levine Conservation Theory for one month at National Referal Hospital in Jakarta. Case studies are carried out by applying the nursing process according to Levine’s Conservation Theory, which consists of trophicognosis, hypothesis, nursing intervention and evaluation. Data analysis within cases and across cases is conducted by content analysis related to Levine’s evaluation model.
 Results: The results of the case study show that eight clients experienced energy conservation imbalances, ten clients experienced structural integrity disorders and eight clients experienced impaired social integrity. The results of the application of the Levine Conservation Theory for one month showed only two clients who could not achieved energy conservation and structural integrity.
 Conclusion/Importance: This case study shows that the Levine conservation model can be used in the nursing process in children with mucositis to maintain energy conservation and structural integrity. Nurses are expected to understand the application of Levine’s conservation theory to clients who are prone to conservation problems.

Prostate cancer cells enhance interleukin-15-mediated expansion of NK cells.

To identify cytokines that can activate and expand NK cells in the presence of prostate cancer cells in order to determine whether these agents may be useful in future intra-tumoural administration in pre-clinical and clinical prostate cancer trials. Lymphocytes isolated from normal donor blood were set up in co-cultures with either cancer or non-cancerous prostate cell lines, together with each of the cytokines interleukin (IL)-2, IL-12, IL-15, interferon (IFN)-γ or IL-21 for a period of 7days. Then, expansion of NK cells, NKT cells and CD8 T cells was measured by flow cytometry and compared with the expansion of the same cells in the absence of prostate cells. The cytotoxic activity of NK cells, as measured by perforin and tumour cell killing, was also assessed. NK cell receptors and their corresponding ligands on prostate tumour cells were analysed to determine whether any of these were modulated by co-culture. The role of the tumour-secreted heat shock proteins HSP90 and HSP70 in the expansion of NK cells in the co-cultures was also investigated because of their effects on NK and CD8 T-cell activation. We showed that, among a panel of cytokines known to cause NK cell activation and expansion, only IL-15 could actively induce expansion of NK, NKT and CD8 T cells in the presence of prostate cancer cell lines. Furthermore, the expansion of NK cells was far greater (up to 50% greater) in the presence of the cancer cells (LNCaP, PC3) than when lymphocytes were incubated alone. In contrast, non-cancerous cell lines (PNT2 and WPMY-1) did not exert any expansion of NK cells. The cytolytic activity of the NK cells, as measured by perforin, CD107a and killing of tumour cells, was also greatest in co-cultures with IL-15. Examination of NK cell receptors shows that NKG2D is upregulated to a greater degree in the presence of prostate cancer cells, compared with the upregulation with IL-15 in lymphocytes alone. However, blocking of NKG2D does not inhibit the enhanced expansion of NK cells in the presence of tumour cells. Among a panel of NK cell-activating cytokines, IL-15 was the only cytokine that could stimulate expansion of NK cells in the presence of prostate cancer cells; therefore IL-15 may be a good candidate for novel future intra-tumoural therapy of the disease.

Abstract 2746: Stat3 labels a subpopulation of reactive astrocytes required for brain metastasis

Abstract The diagnosis of brain metastasis involves high morbidity and mortality and remains as an unmet clinical need in spite of being the most common tumor in the brain. Brain metastasis affects between 10-30% of cancer patients with 200.000 new cases yearly only in the US. Increasing evidences point out the relevance of the microenvironment to understand the biology of brain colonization by metastatic cells, however translation strategies targeting it are lacking. We have identified altered signaling pathways in pro-metastatic reactive astrocytes (RA), and translated that into a novel therapeutic application for brain metastasis. Specifically we have identified a subpopulation of RA characterized by activated STAT3 pathway (pSTAT3+). This subpopulation is located in the vicinity of metastatic lesions intermingled with pSTAT3- RA in several experimental models and in 89% of human brain metastases independently of the primary tumor source. The pro-metastatic behaviour of RA is regulated by the activation of the STAT3 pathway, which modulates the immune system locally promoting the survival of cancer cells. Specifically the secretome of pSTAT3+ RA decreases the anti-tumor activity of CD8+ T cells as well as promotes the expansion of the pro-tumor population of CD74+ microglia/macrophage, which infiltrates metastasis cores. Interestingly, the immunosuppressive nature of pSTAT3+ RA is linked to the acquisition of stem cell-like properties, which might reflect the misuse of brain responses to injury instigated by the presence of cancer cells. Genetic and pharmacologic approaches targeting STAT3 in RA impair the progression of brain metastasis, even at advanced stages of the disease. Moreover, a safe and orally bioavailable STAT3 inhibitor reduced brain metastasis in 75% of 18 stage IV lung adenocarcinoma patients with established brain metastases, improving the outcome of the disease by increasing patient survival from 4 to 15 months. Besides increasing therapeutic opportunities for patients with brain metastasis, we have described for the first time the role of reactive astrocytes as regulators of local immunosuppression in brain metastasis as well as the importance of uncovering the heterogeneity within the metastasis-associated microenvironment. Citation Format: Neibla Priego, Lucía Zhu, Catia Monteiro, Manon Mulders, David Wasilewski, Wendy Bindeman, Laura Doglio, Elena Martínez-Saez, Santiago Ramón y Cajal, Coral Fustero-Torre, Elena Piñeiro-Yáñez, Aurelio Hernández-Laín, Valeria Poli, Javier A. Menéndez, Ricardo Soffietti, Joaquim Bosch-Barrera, Manuel Valiente. Stat3 labels a subpopulation of reactive astrocytes required for brain metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2746.

An Electron paramagnetic resonance (EPR) spin labeling study in HT-29 Colon adenocarcinoma cells after Hypericin-mediated photodynamic therapy

BackgroundColon cancer affects 1.23 million people worldwide and is the third most common malignant disease in men and the second in women. The only curative treatment is surgical resection, but a significant number of patients develop local recurrence or distant metastases. One of the alternative treatment methods for colon cancer is photodynamic therapy (PDT). In recent years, hypericin (HYP) derived from Hypericum perforatum has been suggested as a strong candidate photosensitizer for PDT. Our interest is focused on the biophysical changes in colon cancer cells in relation to HYP-mediated PDT.ResultsIn this study, HYP-mediated PDT at 0.04, 0.08 or 0.15 μM HYP concentrations was performed in HT-29 colon adenocarcinoma cells and the Electron Paramagnetic Resonance (EPR) spectra of the spin labeled cells were obtained. Plasma membranes are already heterogeneous structures; the presence of cancer cells increased the heterogeneity and also fluidity of the plasma membranes. Therefore, the obtained spectra were evaluated by EPRSIMC program, which provides the calculation of heterogeneous structures up to four spectral components with different fluidity characteristics. Generally, two motional patterns were obtained from calculations and the number of them increased at the highest concentration. As the order parameters of the most populated components compared, an increase was observed depending on the HYP concentration. However, because of the heterogeneous structure of membrane, the order parameters of the less populated components did not exhibit a regular distribution.ConclusionAfter HYP-mediated PDT, concentration dependent changes were observed in the domain parameters indicating an increase in the HYP accumulation.

GSK3\u03b2 regulates epithelial-mesenchymal transition and cancer stem cell properties in triple-negative breast cancer

BackgroundTriple-negative breast cancers (TNBCs), which lack receptors for estrogen, progesterone, and amplification of epidermal growth factor receptor 2, are highly aggressive. Consequently, patients diagnosed with TNBCs have reduced overall and disease-free survival rates compared to patients with other subtypes of breast cancer. TNBCs are characterized by the presence of cancer cells with mesenchymal properties, indicating that the epithelial to mesenchymal transition (EMT) plays a major role in the progression of this disease. The EMT program has also been implicated in chemoresistance, tumor recurrence, and induction of cancer stem cell (CSC) properties. Currently, there are no targeted therapies for TNBC, and hence, it is critical to identify the novel targets to treat TNBC.MethodsA library of compounds was screened for their ability to inhibit EMT in cells with mesenchymal phenotype as assessed using the previously described Z-cad reporters. Of the several drugs tested, GSK3β inhibitors were identified as EMT inhibitors. The effects of GSK3β inhibitors on the properties of TNBC cells with a mesenchymal phenotype were assessed using qRT-PCR, flow cytometry, western blot, mammosphere, and migration and cell viability assays. Publicly available datasets also were analyzed to examine if the expression of GSK3β correlates with the overall survival of breast cancer patients.ResultsWe identified a GSK3β inhibitor, BIO, in a drug screen as one of the most potent inhibitors of EMT. BIO and two other GSK3β inhibitors, TWS119 and LiCl, also decreased the expression of mesenchymal markers in several different cell lines with a mesenchymal phenotype. Further, inhibition of GSK3β reduced EMT-related migratory properties of cells with mesenchymal properties. To determine if GSK3β inhibitors target mesenchymal-like cells by affecting the CSC population, we employed mammosphere assays and profiled the stem cell-related cell surface marker CD44+/24− in cells after exposure to GSK3β inhibitors. We found that GSK3β inhibitors indeed decreased the CSC properties of cell types with mesenchymal properties. We treated cells with epithelial and mesenchymal properties with GSK3β inhibitors and found that GSK3β inhibitors selectively kill cells with mesenchymal attributes while sparing cells with epithelial properties. We analyzed patient data to identify genes predictive of poor clinical outcome that could serve as novel therapeutic targets for TNBC. The Wnt signaling pathway is critical to EMT, but among the various factors known to be involved in Wnt signaling, only the higher expression of GSK3β correlated with poorer overall patient survival.ConclusionsTaken together, our data demonstrate that GSK3β is a potential target for TNBCs and suggest that GSK3β inhibitors could serve as selective inhibitors of EMT and CSC properties for the treatment of a subset of aggressive TNBC. GSK3β inhibitors should be tested for use in combination with standard-of-care drugs in preclinical TNBC models.

ATF3 and JDP2 deficiency in cancer associated fibroblasts promotes tumor growth via SDF-1 transcription

The activating transcription factor 3 (ATF3) and the c-Jun dimerization protein 2 (JDP2) are members of the basic leucine zipper (bZIP) family of transcription factors. These proteins share a high degree of homology and both can activate or repress transcription. Deficiency of either one of them in the non-cancer host cells was shown to reduce metastases. As ATF3 and JDP2 compensate each other’s function, we studied the double deficiency of ATF3 and JDP2 in the stromal tumor microenvironment. Here, we show that mice with ATF3 and JDP2 double deficiency (designated thereafter dKO) developed larger tumors with high vascular perfusion and increased cell proliferation rate compared to wild type (WT) mice. We further identify that the underlying mechanism involves tumor associated fibroblasts which secrete high levels of stromal cell-derived factor 1 (SDF-1) in dKO fibroblasts. SDF-1 depletion in dKO fibroblasts dampened tumor growth and blood vessel perfusion. Furthermore, ATF3 and JDP2 were found to regulate SDF-1 transcription and secretion in fibroblasts, a phenomenon that is potentiated in the presence of cancer cells. Collectively, our results suggest that ATF3 and JDP2 regulate the expression of essential tumor promoting factors expressed by fibroblasts within the tumor microenvironment, and thus restrain tumor growth.

Ultrafast Electrochemical Trigger Drug Delivery Mechanism for Nanographene Micromachines

AbstractNano/micromachines with autonomous motion are the frontier of nanotechnology and nanomaterial research. These self‐propelled nano/micromachines convert chemical energy obtained from their surroundings to propulsion. They have shown great potential in diagnostic and therapeutic applications. This work introduces a high‐speed tubular electrically conductive micromachine based on reduced nanographene oxide (n‐rGO) as a platform for drug delivery and platinum (Pt) as the catalytic inner layer. n‐rGO/Pt micromachines are loaded with doxorubicin (DOX) by a simple physical adsorption with a very high loading efficiency, displaying single‐ or multistrand wrapping of DOX monomers on the micromachine cylinders. More importantly, it is found that electron injection into DOX@n‐rGO/Pt micromachines via electrochemistry leads to expulsion of DOX from micromachines in motion within only a few seconds. An in vitro study confirms this efficient release mechanism in the presence of cancerous cells. The unique properties of the n‐rGO/Pt micromotor enable the effective management of DOX release at the tumor site and thus enhances the therapeutic efficiency and reduces the side toxicity toward the healthy tissue. These micromachine drug carriers combine the high loading capacity of conventional carbon‐based drug carriers with a fast and efficient electrochemical drug‐release mechanism.

The characterization of “islet invasion” in pancreatic ductal adenocarcinoma

Introduction: Insulin and insulin-like growth factor (IGF) signaling axis prompts pancreatic ductal adenocarcinoma/PDAC into an advanced stage by enhancing tumor growth,metastasis and therapy resistance. In the pancreas tissue, the main source of insulin is the Langerhans' islets. Importantly, PDAC cells also invade islets during carcinogenesis and distort their structure, but this “islet invasion” has never been systematically characterized. Method: To classify the islet invasion and structure in PDAC, pancreatic tissues from 75 PDAC patients, with or without diabetes mellitus, have been double-immunostained with the ductal and cancer cell marker Cytokeratin 19 and with insulin. Accordingly, pancreatic islet structure and invasion pattern has been characterized. Results: PDAC tissues showed five different pancreatic “islet invasion patterns”: 1) Presence of cancer cells around the borders of intact pancreatic islets termed ‘peri-insular invasion’, 2) presence of cancer cells within pancreatic islets termed ‘endo-insular invasion’, 3) Distortion of the pancreatic islet structure due to destructive growth of cancer cells (‘distorted islet structure’), 4) Insulin-expressing CK19-positive cancer cells', 5) adjacence of small clusters of endocrine cells to cancer (‘adjacent cancer & islet’). Moreover, we found that a portion of insulin-expressing cells co-expressed mutant K-Ras(G12D). Conclusion: PDAC tissues frequently harbour “islet invasion” and, in addition, insulin expression in KrasG12D-mutated cells. Due to the pivotal role of insulin- and IGF-1-signaling in PDAC, it is essential to elucidate the impact of islet invasion on the pathogenesis and progression of PDAC.

Positive Surgical Margins Predict Progression-free Survival After Nephron-sparing Surgery for Renal Cell Carcinoma: Results From a Single Center Cohort of 459 Cases With a Minimum Follow-up of 5 Years

The role of positive surgical margins (PSMs) on the recurrence of renal cell carcinoma (RCC) after partial nephrectomy (PN) is debated, and available evidence lacks long-term data. The aim of this study was to evaluate the predictive role of PSMs on progression-free survival (PFS) in a large cohort followed for at least 5 years. This study was a retrospective analysis of a prospectively compiled single-institution database collecting complete information on more than 2700 patients who had undergone surgery for renal tumor. The data of all the patients submitted to PN for RCC and with least 5 years follow-up were extracted. Surgical specimens were examined at the time of surgery only by 2 expert uro-pathologists. A PSM was defined as the presence of cancer cells at the inked surface of the specimen. The role of PSMs on survival was estimated by Cox regression models adjusted for influent covariates. A total of 459 patients fulfilled the inclusion criteria and were evaluated. PSMs were observed in 27 (5.9%) cases. No differences in preoperative and pathologic data were found comparing patients with and without PSMs. At a median follow-up of 96 months (interquartile range, 74-131 months), a clinically evident relapse of RCC was diagnosed in 36 (7.8%) patients at a median interval of 36 months from PN. Among these, 6 had a PSM for an incidence of relapse of 22.2% in the PSM group, whereas 30 had negative margins, for an incidence of 6.9% (P=.013). The sites of relapse were distant organs in 18 cases, and the kidney underwent PN in 21. The patients with PSMs showed a borderline significantly higher incidence of distant metastasis (11.1% vs. 3.5%; P= .071) and a significantly higher incidence of renal relapses (14.8% vs. 3.9%; P= .029). Multivariable Cox models confirmed that the presence of PSMs was an independent predictor of PFS (odds ratio, 3.127; P= .013). PSMs are an independent predictor of PFS in patients who underwent PN for RCC, owing to a higher incidence of distant and local relapses. Surveillance in presence of PSMs should be intensified and extended for a long time.

Primary Pulmonary Colloid Adenocarcinoma: How Can We Obtain a Precise Diagnosis?

A 76-year-old asymptomatic man was found to have a mass in the right lower lung field. Although the presence of a mucinous component in the majority of the tumor was shown by magnetic resonance imaging, the presence of cancer cells was suspected by contrast enhancement on computed tomography (CT) and based on the partial accumulation in the marginal regions of the tumor on fluorodeoxyglucose-positron emission tomography (FDG-PET). A transbronchial lung biopsy was non-diagnostic, but resection of the mass resulted in a diagnosis of colloid adenocarcinoma. The findings from combined contrast CT and FDG-PET may raise the suspicion of colloid adenocarcinoma and prompt the consideration of surgical resection.

Myoepithelial cells are a dynamic barrier to epithelial dissemination.

The mammary epithelium is composed of an inner luminal and surrounding myoepithelial cell layer. The presence of cancer cells beyond the myoepithelium defines invasive breast cancer, yet the role of the myoepithelium during invasion remains unclear. We developed a 3D organotypic culture assay to model this process through lineage-specific expression of the prometastatic transcription factor Twist1 We sought to distinguish the functional role of the myoepithelium in regulating invasion and local dissemination. Myoepithelial-specific Twist1 expression induced cell-autonomous myoepithelial cell escape. Remarkably, luminal-specific Twist1 expression was rarely sufficient for escape. Time-lapse microscopy revealed that myoepithelial cells collectively restrain and reinternalize invading Twist1+ luminal cells. Barrier function correlated with myoepithelial abundance and required the expression of α-smooth muscle actin and P-cadherin. We next demonstrated that myoepithelial cells can restrain and recapture invasive cancer cells. Our data establish the concept of the myoepithelium as a dynamic barrier to luminal dissemination and implicate both smooth muscle contractility and intercellular adhesion in barrier function.

WNK1 kinase and its partners Akt, SGK1 and NBC-family Na+/HCO3- cotransporters are potential therapeutic targets for glioblastoma stem-like cells linked to Bisacodyl signaling.

Glioblastoma is a highly heterogeneous brain tumor. The presence of cancer cells with stem-like and tumor initiation/propagation properties contributes to poor prognosis. Glioblastoma cancer stem-like cells (GSC) reside in hypoxic and acidic niches favoring cell quiescence and drug resistance. A high throughput screening recently identified the laxative Bisacodyl as a cytotoxic compound targeting quiescent GSC placed in acidic microenvironments. Bisacodyl activity requires its hydrolysis into DDPM, its pharmacologically active derivative. Bisacodyl was further shown to induce tumor shrinking and increase survival in in vivo glioblastoma models. Here we explored the cellular mechanism underlying Bisacodyl cytotoxic effects using quiescent GSC in an acidic microenvironment and GSC-derived 3D macro-spheres. These spheres mimic many aspects of glioblastoma tumors in vivo, including hypoxic/acidic areas containing quiescent cells. Phosphokinase protein arrays combined with pharmacological and genetic modulation of signaling pathways point to the WNK1 serine/threonine protein kinase as a mediator of Bisacodyl cytotoxic effect in both cell models. WNK1 partners including the Akt and SGK1 protein kinases and NBC-family Na+/HCO3− cotransporters were shown to participate in the compound’s effect on GSC. Overall, our findings uncover novel potential therapeutic targets for combatting glioblastoma which is presently an incurable disease.

Bifunctional aptamer-mediated catalytic hairpin assembly for the sensitive and homogenous detection of rare cancer cells

The presence of cancer cells in body fluids confirms the occurrence of metastasis and guides treatment. A simple, fast, and homogeneous fluorescent method was developed to detect cancer cells based on catalytic hairpin assembly (CHA) and bifunctional aptamers. The bifunctional aptamer had a recognition domain for binding to target cancer cells and an initiator domain for triggering the CHA reaction. In the presence of target cells, the bifunctional aptamer was released from the inhibitor and initiated a cascade reaction of assembly and disassembly of the hairpins. Separation of the fluorophores from the quenchers produced fluorescence signals. The proposed strategy showed high specificity for discriminating normal cells and leukocytes, and the detection limit was 10 cells/mL, which was lower than that of previous aptasensors. This assay was further tested using four kinds of clinical samples spiked with target cells to confirm its applicability. We developed a simple, rapid, and cost-effective method for the detection of cancer cells that did not require purification, and the approach holds great potential for bioanalysis and early diagnosis.

A Phenotypic Cell-Binding Screen Identifies a Novel Compound Targeting Triple-Negative Breast Cancer.

We describe a "phenotypic cell-binding screen" by which therapeutic candidate targeting cancer cells of a particular phenotype can be isolated without knowledge of drug targets. Chemical library beads are incubated with cancer cells of the phenotype of interest in the presence of cancer cells lacking the phenotype of interest, and then the beads bound to only cancer cells of the phenotype of interest are selected as hits. We have applied this screening strategy in discovering a novel compound (LC129-8) targeting triple-negative breast cancer (TNBC). LC129-8 displayed highly specific binding to TNBC in cancer cell lines and patient-derived tumor tissues. LC129-8 exerted anti-TNBC activity by inducing apoptosis, inhibiting proliferation, reversing epithelial-mesenchymal transition, downregulating cancer stem cell activity and blocking in vivo tumor growth.

The interacting role of physical stiffness and tumor cells on the macrophages polarization

There is a complex interplay existing between the tumor microenvironment and cellular function, while the influence of tumor biophysical signals on the macrophage function remain unclear. Herein, we constructed a three dimensional hydrogel model to study the symphysial effect of physical signal (stiffness) and biological signal (tumor cells) on macrophage polarization in different stage of tumor progression. In the absence of biological signal, the macrophage response was slightly changed with different stiffness (100 Pa, 300 Pa, and 900 Pa). Once co-incubated with the tumor cell, macrophage induced more significant cancer facilitated cytokines (e.g. IL-10, MMP-9) and chemokines (MCP-1). In contrast to the education role from the co-cultured cancer cells, a stiffness around the normal tissue (100 Pa, CO-L group) provided a physical cue for cancer cell regression. This cue triggered the active role of M1 type macrophages (e.g. IL-12), indicating the interacting role of stiffness in the presence of cancer cell. The stiffness around the premalignant tissue (M, 300 Pa) induced much higher level of cytokines (e.g. IL-6, IL-10, IL-12, CD86, and CD206), while the immune activation cytokines (M1 type) was largely attenuated at high stiffness (900 Pa). This study revealed the pleiotropic tuning of macrophage function under different biophysical signals, which provided important knowledge for cancer progress and may benefit the design of cancer-related therapies.

Volatile metabolomic signature of bladder cancer cell lines based on gas chromatography–mass spectrometry

Recent studies provide a convincing support that the presence of cancer cells in the body leads to the alteration of volatile organic compounds (VOCs) emanating from biological samples, particularly of those closely related with tumoral tissues. Thus, a great interest emerged for the study of cancer volatilome and subsequent attempts to confirm VOCs as potential diagnostic biomarkers. The aim of this study was to determine the volatile metabolomic signature of bladder cancer (BC) cell lines and provide an in vitro proof-of-principle that VOCs emanated into the extracellular medium may discriminate BC cells from normal bladder epithelial cells. VOCs in the culture media of three BC cell lines (Scaber, J82, 5637) and one normal bladder cell line (SV-HUC-1) were extracted by headspace-solid phase microextraction and analysed by gas chromatography-mass spectrometry (HS-SPME/GC-MS). Two different pH (pH 2 and 7) were used for VOCs extraction to infer the best pH to be used in in vitro metabolomic studies. Multivariate analysis revealed a panel of volatile metabolites that discriminated cancerous from normal bladder cells, at both pHs, although a higher number of discriminative VOCs was obtained at neutral pH. Most of the altered metabolites were ketones and alkanes, which were generally increased in BC compared to normal cells, and alcohols, which were significantly decreased in BC cells. Among them, three metabolites, namely 2-pentadecanone, dodecanal and γ-dodecalactone (the latter only tentatively identified), stood out as particularly important metabolites and promising volatile biomarkers for BC detection. Furthermore, our results also showed the potential of VOCs in discriminating BC cell lines according to tumour grade and histological subtype. We demonstrate that a GC-MS metabolomics-based approach for analysis of VOCs is a valuable strategy for identifying new and specific biomarkers that may improve BC diagnosis. Future studies should entail the validation of volatile signature found for BC cell lines in biofluids from BC patients.

Risk of transplanting malignant cells in cryopreserved ovarian tissue.

Improvements in cancer treatments have increased the chances of survival of young cancer patients, but have given rise to other issues, like premature ovarian insufficiency and infertility. Preservation and subsequent restoration of ovarian function in these patients is now possible thanks to ovarian tissue cryopreservation and transplantation. However, safety concerns about the possible presence of cancerous cells in the tissue to be cryopreserved must be addressed. Indeed, reimplantation of malignant cell-contaminated ovarian tissue could potentially lead to recurrence of the primary disease. This review of the existing literature aims to evaluate the risk of reintroducing malignant cells in pathologies that represent the main indications for ovarian tissue cryopreservation. Experimental studies applying molecular analyses as well as xenografting investigation have proved that hematologic and ovarian malignancies are those at greatest risk of ovarian involvement, and hence recurrence upon reimplantation of frozen-thawed ovarian tissue. Preimplantation analysis using highly sensitive techniques and disease-specific markers is therefore vital in these patients to detect minimal disseminate disease.

MP48-02 POSITIVE SURGICAL MARGINS PREDICT PROGRESSION-FREE SURVIVAL AFTER NEPHRON SPARING SURGERY FOR RENAL CELL CARCINOMA: RESULTS FROM A SINGLE CENTRE COHORT OF ALMOST 500 CASES WITH A MINIMUM FOLLOW-UP OF 5 YEARS

You have accessJournal of UrologyKidney Cancer: Localized: Surgical Therapy V1 Apr 2018MP48-02 POSITIVE SURGICAL MARGINS PREDICT PROGRESSION-FREE SURVIVAL AFTER NEPHRON SPARING SURGERY FOR RENAL CELL CARCINOMA: RESULTS FROM A SINGLE CENTRE COHORT OF ALMOST 500 CASES WITH A MINIMUM FOLLOW-UP OF 5 YEARS Riccardo Tellini, Alessandro Antonelli, Carlotta Palumbo, Alessandro Veccia, Maria Furlan, Regina Tardanico, Simona Fisogni, and Claudio Simeone Riccardo TelliniRiccardo Tellini More articles by this author , Alessandro AntonelliAlessandro Antonelli More articles by this author , Carlotta PalumboCarlotta Palumbo More articles by this author , Alessandro VecciaAlessandro Veccia More articles by this author , Maria FurlanMaria Furlan More articles by this author , Regina TardanicoRegina Tardanico More articles by this author , Simona FisogniSimona Fisogni More articles by this author , and Claudio SimeoneClaudio Simeone More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1501AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The real role of positive surgical margins (PSM) after nephron sparing surgery (NSS) on progression and relapse of disease is still debated because strong evidences are lacking, especially at a long term horizon. The aim of this study was to evaluate the oncologic impact of PSM in a large contemporary single centre cohort of PN followed for a long time. METHODS A retrospective analysis of our prospectively maintained institutional database on renal surgery since 1983 was performed. The indication to radical versus partial nephrectomy (PN) followed the contemporary international guidelines; the surgical strategy for PN was generally an enucleoresection. All surgical specimens were examined by two expert uro-pathologists. For the present study, all patients who underwent PN for non metastatic RCC with at least 5 years of follow-up, if uneventful, were included; the patients died from non cancer causes within the minimum time of follow-up required were excluded. A PSM was defined as the presence of cancer cells at the inked surface of the final specimen. The primary endpoint was progression free survival (PFS), considering as events of progression all recurrences of disease at any site. A Cox regression model estimated the role of PSM on survival, after adjustment for influent covariates. RESULTS Out of 490 patients submitted to NSS for RCC, 459 patients fulfilled inclusion criteria and were evaluated for the purpose of this study. PSM were observed in 27 (5.9%) cases. No differences in terms of preoperative and pathological features of patients with positive and negative surgical margins (NSM) were found. At a mean follow-up of 110 (±51.9) months, progression was diagnosed in 36 (7.8%) patients overall. Among these patients, 6 had PSM (6 out 27, 22.2%) and 30 (out of 432, 6.9%) had NSM (p= 0.013). The sites of progression were distant organs in 18 cases and the operated kidney in 21. No differences were found in terms of incidence of distant metastasis between PSM and NSM groups, whereas there was a significant difference in the incidence of relapses on the operated kidney (14.8% versus 3.9%; p= 0.029). At multivariable Cox regression analysis, a PSM was confirmed as an independent predictor of survival [OR: 3.127 (IQR 1.272 – 7.688); p=0.013]. CONCLUSIONS A PSM is an independent predictor of PFS in patients submitted to NSS for RCC and followed for a long time, mainly due to a higher rate of local relapses of disease. In these patients, a tailored follow-up is mandatory. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e624 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Riccardo Tellini More articles by this author Alessandro Antonelli More articles by this author Carlotta Palumbo More articles by this author Alessandro Veccia More articles by this author Maria Furlan More articles by this author Regina Tardanico More articles by this author Simona Fisogni More articles by this author Claudio Simeone More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Abstract A57: Glutathione depletion overcomes chemotherapy resistance in aggressive medulloblastoma stem-like cells

Abstract Drug resistance is one of the main limiting steps for the successful treatment of most cancers. Additionally, the presence of cancer cells with stem cell traits favors the process of treatment resistance, tumor recurrence, and spreading. Glutathione (GSH) is a major antioxidant intracellular molecule and has been positively correlated with the development of drug resistance. The purpose of this study was to verify whether GSH depletion potentiates the effect of chemotherapy in stem-like cells from the medulloblastoma cell line Daoy. Cells expressing normal (control tumor cells) or high levels of the pluripotency factor OCT4A (stem-like cells) were treated with a combination of BSO (glutathione inhibitor), cisplatin, and temozolomide (TMZ). Cell viability was assessed through MTT and apoptosis assays. Initially, the concentration of BSO required to deplete the GSH in both Daoy and Daoy-OCT4A cells was established (50 and 58 μM, respectively), as well as the IC50 of each chemotherapeutic drug. Both cells were sensitive to cisplatin (IC50 = 0.494 μM for Daoy and 0.215 μM for Daoy-OCT4A), but resistant to TMZ. The highest dose of TMZ used (500 μM) reduced the viability of Daoy cells to 56%, but only to 80% in Daoy-OCT4A cells, denoting the chemoresistant phenotype of these cancer stem-like cells. Next, the ideal combination of the three compounds was defined. For Daoy cells, it was BSO 50 μM + cisplatin 0.8 μM + TMZ 0.8 μM. As for Daoy-OCT4A cells, it was BSO 58 μM + cisplatin 0.215 μM + TMZ 500 μM. Daoy-OCT4A cells required a higher dose of TMZ than control Daoy cells to attain similar death rates; however, the combined treatment was effective for both cells. When combining these three drugs, a synergistic effect was observed for Daoy-OCT4A cells, reducing cell viability to ~17%, in comparison with treatment of cisplatin+TMZ alone. As for Daoy cells, an additive effect was observed and viability was reduced to ~15%, in comparison with cisplatin+TMZ alone. This indicates that the use of a glutathione inhibitor in combination with cisplatin and TMZ may be useful in the treatment of medulloblastoma, even in the case of aggressive tumors. Similarly, this approach may also be useful in the development of therapeutic strategies for other chemoresistant tumors. Citation Format: Beatriz Dias Barbieri, Marina Marçola, Clarissa Rocha, Oswaldo Keith Okamoto. Glutathione depletion overcomes chemotherapy resistance in aggressive medulloblastoma stem-like cells [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A57.