Interactions between the tumor and the blood systemic response of breast cancer patients.

Although systemic immunity is critical to the process of tumor rejection, cancer research has largely focused on immune cells in the tumor microenvironment. To understand molecular changes in the patient systemic response (SR) to the presence of BC, we profiled RNA in blood and matched tumor from 173 patients. We designed a system (MIxT, Matched Interactions Across Tissues) to systematically explore and link molecular processes expressed in each tissue. MIxT confirmed that processes active in the patient SR are especially relevant to BC immunogenicity. The nature of interactions across tissues (i.e. which biological processes are associated and their patterns of expression) varies highly with tumor subtype. For example, aspects of the immune SR are underexpressed proportionally to the level of expression of defined molecular processes specific to basal tumors. The catalog of subtype-specific interactions across tissues from BC patients provides promising new ways to tackle or monitor the disease by exploiting the patient SR.

Peripheral blood cells inform on the presence of breast cancer: a population-based case-control study.

Tumor-host interactions extend beyond the local microenvironment and cancer development largely depends on the ability of malignant cells to hijack and exploit the normal physiological processes of the host. Here, we established that many genes within peripheral blood cells show differential expression when an untreated breast cancer (BC) is present, and harnessed this fact to construct a 50-gene signature that distinguish BC patients from population-based controls. Our results were derived from a series of large datasets within our unique population-based Norwegian Women and Cancer cohort that allowed us to investigate the influence of medications and tumor characteristics on our blood-based test, and were further tested in two external datasets. Our 50-gene signature contained cytostatic signals including the specific suppression of the immune response and medications influencing transcription involved in those processes were identified as confounders. Through analysis of the biological processes differentially expressed in blood, we were able to provide a rationale as to why the systemic response of the host may be a reliable marker of BC, characterized by the underexpression of both immune-specific pathways and "universal" cell programs driven by MYC (i.e., metabolism, growth and cell cycle). In conclusion, gene expression of peripheral blood cells is markedly perturbed by the specific presence of carcinoma in the breast and these changes simultaneously engage a number of systemic cytostatic signals emerging connections with immune escape of BC.

Detection of DNA double-strand breaks using gammaH2AX after MRI exposure at 3 Tesla: an in vitro study.

PURPOSE: To evaluate the effects of the static magnetic field and typical imaging sequences of a high-field magnetic resonance scanner (3 Tesla) on the induction of double-strand breaks (DSBs) in two different human cell lines. MATERIALS AND METHODS: Human promyelocytic leukemia cells (HL-60) and human acute myeloid leukemia cells (KG-1a) were exposed to the static magnetic field alone and to turbo spin-echo (TSE) and gradient-echo (GE) sequences. Flow cytometry was used to quantify gammaH2AX (serine 139 phosphorylated form of histone H2AX) expression of antibody-stained cells as a marker for deoxyribonucleic acid (DNA) DSBs one hour and 24 hours after magnetic field exposure. X-ray-treated cells were used as positive control. RESULTS: Neither exposure to the static magnetic field alone nor to the applied imaging sequences showed significant differences in gammaH2AX expression between exposed and sham-exposed cells. X-ray-treated cells as positive control showed a significant increase in gammaH2AX expression. CONCLUSION: The static magnetic field alone and MRI sequences at 3 Tesla have no effect on the induction of DSBs in HL-60 and KG-1a cells.

Do static or time-varying magnetic fields in magnetic resonance imaging (3.0 T) alter protein-gene expression?-A study on human embryonic lung fibroblasts.

PURPOSE: To evaluate the influence of magnetic resonance imaging (MRI) on gene expression in embryonic human lung fibroblasts (Hel 299). MATERIALS AND METHODS: The cells were exposed to the static magnetic field and to a turbo spin-echo sequence of an MR scanner at 3.0 Tesla. An MR group (exposed) and a control group (sham-exposed) were set up using a special MR-compatible incubation system. The exposure time was two hours. Gene expression profiles were studied using a complementary deoxyribonucleic acid (cDNA) microarray containing 498 known genes involved in transcription, intracellular transport, structure/junction/adhesion or extracellular matrix, signaling, host defense, energetics, metabolism, cell shape, and death. RESULTS: No changes in gene expression were found in either group (exposed or sham-exposed cells) at the end of a two-hour exposure for any of the 498 tested protein genes. CONCLUSION: The results suggest that MRI has no influence on protein-gene expression in eugenic human lung cells.

In vitro evaluation of magnetic resonance imaging at 3.0 tesla on clonogenic ability, proliferation, and cell cycle in human embryonic lung fibroblasts.

OBJECTIVES: We investigated the influence of magnetic resonance (MR) at 3.0 T on clonogenic ability, proliferation, and cell cycle in an embryonic human cell line. MATERIALS AND METHODS: Cells (human lung fibroblasts Hel 299) were exposed to the static magnetic field (3.0 T) of a magnetic resonance imager (MRI) and to a turbo spin echo sequence at 3.0 T within clinical limitations (specific absorption rate 0.92 W/kg). A special MR-compatible incubation system was used. A control group (sham-exposed) and a MRI group (exposed) were set up. We investigated 3 biologic endpoints: colony forming, cell cycle, and proliferation ability. The exposure time was 2 hours in each experiment. RESULTS: In the statistical analysis, none of these tests showed relevant differences between the exposed and sham-exposed group. CONCLUSIONS: No influences of the static field alone as well as a turbo spin echo sequence at 3.0 T on clonogenic ability, proliferation, or cell cycle in eugenic human lung fibroblasts were found.

MRI-compatible incubation chamber for cell culture experiments.

PURPOSE: To develop an incubation chamber that is compatible with MRI, while avoiding sources of error due to the experimental setup. MATERIALS AND METHODS: Two identical and gas-tight chambers were constructed of Plexiglas. The temperature and the CO(2) concentration were adjustable. Temperature variations within and between both chambers were measured. The pH values of the cell culture media were measured under different environmental settings. For each environment a colony formation test was carried out. The homogeneity of the magnetic field inside the chambers was estimated by phantom tests. RESULTS: The temperature variations within the chambers were <0.3 degrees C, and the variation between the chambers was on average <0.05 degrees C. After eight hours the pH values of the cell culture media were 7.47 and 7.48 in the reference and measurement chambers, respectively; 7.41 in the CO(2) incubator; and 8.73 in ambient air. In colony formation tests the survival fraction in the Plexiglas chamber was comparable to that in the CO(2) incubator. No distortions of the magnetic field from the incubation chamber were observed. CONCLUSION: The incubation system presented can provide the conditions of a CO(2) incubator without alteration of the magnetic flux density.

Subcapsular liver hematoma in HELLP syndrome: Evaluation of diagnostic and therapeutic options--a unicenter study.

OBJECTIVE: Subcapsular liver hematoma formation has been reported in less than 2% of pregnancies complicated by HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome. The purpose of this study was to identify the main diagnostic and therapeutic options for management of these patients. STUDY DESIGN: In this 10-year retrospective review, we performed a computer-directed search of all cases of confirmed HELLP syndrome with hepatic hematoma treated in the surgical department of our tertiary care referral medical center. RESULTS: Five patients with subcapsular liver hematoma in HELLP syndrome could be identified. All patients received transabdominal ultrasound, computed tomography, or magnetic resonance imaging. Conservative treatment was successful in three patients. Emergency surgical intervention was necessary in two patients, including one liver transplantation. CONCLUSION: The case series shows the full diagnostic spectrum with transabdominal ultrasound, computed tomography, and magnetic resonance imaging, as well as the different therapeutic options varying from conservative therapy to operative management, including liver transplantation.

Impact of glafenine hydrochloride on human endothelial cells and human vascular smooth muscle cells: a substance reducing proliferation, migration and extracellular matrix synthesis.

The aim of this study was to examine the effects of glafenine hydrochloride (a nonsteroidal anti-inflammatory drug) on proliferation, clonogenic activity, cell-cycle, migration, and the extracellular matrix protein tenascin of human aortic smooth muscle cells (haSMCs) and human endothelial cells (ECs) in vitro.HaSMCs and ECs were seeded in tissue culture flasks. The cells were treated for 4 days with glafenine hydrochloride (10 microM, 50 microM, 100 microM). Half of the treated groups were incubated again with glafenine hydrochloride, the other half received medium free of glafenine hydrochloride every 4 days until day 20. The growth kinetics and clonogenic activity were assessed. Cell cycle distribution was investigated by FACS, migratory ability was evaluated, and effects on extracellular matrix synthesis were assessed by immunofluorescence. Glafenine hydrochloride inhibited the proliferation and clonogenic activity of haSMCs and ECs in a dose-dependent manner. A block in the G2/M phase and a reduction in the G1 phase occurred. The migratory ability of haSMCs was impaired in a dose-dependent manner and the extracellular matrix protein tenascin was reduced. As glafenine hydrochloride has the ability to fully inhibit proliferation and to partially inhibit migration in haSMCs, it could be an interesting substance for further research in the field of restenosis therapy.

Antiproliferative effects of the antiallergic agent azelastine on human aortic smooth-muscle cells: an in vitro study.

RATIONALE AND OBJECTIVES: The aim of the study was to examine the effects of azelastine on proliferation, clonogenic activity, cell-cycle, and migration of human aortic smooth-muscle cells (haSMCs) in vitro. METHODS: HaSMCs were treated for 4 days with azelastine (1 micromol/L, 25 micromol/L, 50 micromol/L). Half of the treated groups were incubated again with azelastine, the other half received azelastine-free medium every 4 days until day 20. The growth kinetics and clonogenic activity were assessed. The cell-cycle distribution was investigated by FACS -- analysis and the migratory ability was evaluated. RESULTS: Azelastine inhibited the proliferation and the clonogenic activity of haSMCs in a dose dependent manner. A G2/M-phase block was induced and the migratory ability was significantly impaired. CONCLUSION: Azelastine has the potential to inhibit the proliferation of haSMCs. If a sufficient dose can be applied either systemically or locally it could be a valuable substance to prevent restenosis.

Meclofenamic acid for inhibition of human vascular smooth muscle cell proliferation and migration: an in vitro study.

PURPOSE: The aim of the study was to examine the effects of meclofenamic acid on proliferation, clonogenic activity, migratory ability, cell cycle distribution and p44/42 MAPK (mitogen activated protein kinase) expression in serum-stimulated human aortic smooth muscle cells (haSMCs). METHODS: haSMCs were treated with meclofenamic acid in three different concentrations (10 mM, 100 mM, 200 mM) for 4 days. Then meclofenamic acid-free culture medium was supplemented until day 20. Growth kinetics were assessed. Cell cycle analysis was performed by flow cytometry. Clonogenic activity was evaluated with colony formation assays. Migratory ability was investigated by stimulation with platelet-derived growth factor (PDGF-BB) in 24-well plates with 8 mm pores membrane inserts. p44/42 MAPK was detected by Western blot technique. RESULTS: Meclofenamic acid inhibited the proliferation, clonogenic activity and migratory ability of haSMCs in a dose-dependent manner. Cell cycle analysis revealed a G2/M-phase block. The p44/42 MAPK was significantly reduced. CONCLUSION: Meclofenamic acid inhibits the proliferation and migration of haSMCs. If a sufficient dose of meclofenamic acid can be applied systemically or by local drug delivery it could be a valuable substance to prevent restenosis after angioplasty.

Flufenamic acid: growth modulating effects on human aortic smooth muscle cells in vitro.

PURPOSE: The aim of the study was to examine the effects of flufenamic acid on proliferation, clonogenic activity, migratory ability, cell-cycle distribution, and p44/42-mitogen-activated protein kinase (MAPK) expression on serum-stimulated human aortic smooth muscle cells (haSMCs) in vitro. MATERIALS AND METHODS: HaSMCs were treated with flufenamic acid in three different doses (40 micromol/L, 200 micromol/L, 400 micromol/L) for 4 days, and then flufenamic-acid-free culture medium was supplemented every 4 days until day 20 after initial treatment. The growth kinetics were assessed. Cell-cycle analysis was performed by flow cytometry. The clonogenic activity was evaluated with use of colony formation assays. The migratory ability was investigated by stimulation with platelet derived growth factor (PDGF-BB) in 24 well plates with 8-microm pore membrane inserts. The p44/42 MAPK was detected by Western blot technique. RESULTS: Flufenamic acid inhibited the proliferation (400 micromol/L treatment over 4 d; 179,700 +/- 49,800 vs 747,900 +/- 144,000; P <.001), clonogenic activity (400 micromol/L treatment over 4 d; 1 +/- 0.3 vs 50 +/- 1.4; P <.001) and migratory ability (400 micromol/L treatment over 4 d; 8 cells +/- 2 vs 48 cells +/- 15; P <.001) of haSMCs in a dose-dependent manner. Cell-cycle analysis revealed a G2/M-phase block (400 micromol/L treatment over 4 d; 28.9 +/- 1.5 vs 9.5 +/- 3.2; P <.001). The expression of p44/42 MAPK was reduced for a treatment with 400 micromol/L flufenamic acid (controls, 427 BLU +/- 0.305 vs treatment group, 190 BLU +/- 106; P <.05) CONCLUSION: Flufenamic acid inhibits the proliferation and migration of haSMCs. Further experiments with animal models concerning stenosis and restenosis are necessary to evaluate the potential of this promising drug.