The inhibitor of kappa B kinase-epsilon regulates MMP-3 expression levels and can promote lung metastasis.

The factors that determine the ability of metastatic tumor cells to expand and grow in specific secondary site(s) are not yet fully understood. Matrix metalloproteinases (MMP) were identified as potential regulators of the site-specificity of metastasis. We found that lung carcinoma cells ectopically expressing high levels of the receptor for the type I insulin like growth factor receptor (M27(R) cells) had a significant reduction in MMP-3 expression levels and this coincided with reduced metastasis to the lung. We used these cells to further investigate signaling pathways regulating MMP-3 expression and the role that MMP-3 plays in lung metastasis. We show that ectopic IκB kinase ɛ (IKKɛ) expression in these cells partly restored MMP-3 expression levels and also sensitized MMP-3 transcription to induction by phorbol 12-myristate 13-acetate (PMA). This increase in MMP-3 production was due to increased activation of several signal transduction mediators, including protein kinase C alpha, ERK2, Akt and the transcription factor p65. Furthermore, reconstitution of MMP-3 expression in M27(R) cells restored their ability to colonize the lung whereas silencing of MMP-3 in M27 cells reduced metastases. Collectively, our results implicate IKKɛ as a central regulator of PMA-induced cell signaling and MMP-3 expression and identify MMP-3 as an enabler of tumor cell expansion in the lung.Oncogenesis (2014) 3, e116; doi:10.1038/oncsis.2014.28; published online 18 August 2014.

Sustained production of a soluble IGF-I receptor by gutless adenovirus-transduced host cells protects from tumor growth in the liver.

The IGF-I receptor (IGF-IR) has an important role in malignant disease and is the target of several drugs presently in clinical trials. Gene therapy has been explored as cancer treatment, mainly for delivery of genes that induce cell death or enhance the immunological response to cancer. Previously, we have shown that the implantation of autologous bone-marrow stromal cells producing a soluble form of IGF-IR (sIGFIR) inhibited experimental liver metastasis of several tumor types in mice. Here, we evaluated the utility of adenovirus-based gene delivery for generating therapeutically effective plasma levels of this decoy. We constructed a third generation gutless adenovirus expressing sIGFIR and found that HEK-293 cells transduced by this, but not control adenoviruses, secreted soluble receptor protein that blocked IGF-I-induced tumor cell migration, proliferation and survival in vitro. Following virus injection in vivo, viral DNA was detectable by PCR in several host organs, particularly the liver, and this resulted in the production of measurable sIGFIR plasma levels for up to 21 days post injection. In mice producing virus-encoded sIGFIR, experimental liver metastasis was inhibited, indicating that sIGFIR levels were therapeutically effective. The results show that adenovirus-based delivery of inhibitory soluble proteins can provide an effective anticancer strategy.

Downregulation of cell surface CA125/MUC16 induces epithelial-to-mesenchymal transition and restores EGFR signalling in NIH:OVCAR3 ovarian carcinoma cells.

BACKGROUND: Epithelial ovarian cancer (EOC) cells are prone to metastasise throughout the peritoneal cavity. The epithelial-to-mesenchymal transition (EMT) is a necessary step towards metastatic tumour progression. CA125/MUC16 mucin is a high-molecular-weight glycoprotein overexpressed in the majority of serous carcinomas, suggesting a possible role in the pathogenesis of these cancers. METHODS: The role of CA125/MUC16 in EMT was investigated using single-chain antibody-mediated knockdown of cell surface CA125/MUC16 in overexpressing EOC NIH:OVCAR3 cells. RESULTS: CA125/MUC16 knockdown was associated with morphological alterations along with decreased surface expression of epithelial markers (E-cadherin, cytokeratin-18) and increased expression of mesenchymal markers (N-cadherin, vimentin). Co-immunoprecipitation experiments revealed that CA125/MUC16 binds to E-cadherin and ß-catenin complexes. The in vitro studies showed disruption of cell-cell junctions, enhanced motility, migration and invasiveness in CA125/MUC16 knockdown cells. Enhanced epidermal growth factor receptor (EGFR) activation was observed in CA125/MUC16 knockdown cells along with increased Akt and ERK1/2 phosphorylation, which are downstream effectors of EGFR, and increased MMP-2 and MMP-9 expression and activities. Epidermal growth factor receptor inhibition strongly inhibited the motility of CA125/MUC16 knockdown cells. CONCLUSIONS: Our findings suggest that CA125/MUC16 plays a role in EMT, presumably through its interaction with E-cadherin and ß-catenin complexes and by modulating EGFR and its downstream signalling pathway in NIH:OVCAR3 cells.

Reversible quantum interface for tunable single-sideband modulation.

Using electromagnetically induced transparency in a cesium vapor, we demonstrate experimentally that the quantum state of a light beam can be mapped into the long-lived Zeeman coherences of an atomic ground state. Two noncommuting variables carried by light are simultaneously stored and subsequently read out, with no noise added. We compare the case where a tunable single sideband is stored independently of the other one to the case where the two symmetrical sidebands are stored using the same electromagnetically induced transparency window.

Radiation-pressure cooling and optomechanical instability of a micromirror.

Recent table-top optical interferometry experiments and advances in gravitational-wave detectors have demonstrated the capability of optical interferometry to detect displacements with high sensitivity. Operation at higher powers will be crucial for further sensitivity enhancement, but dynamical effects caused by radiation pressure on the interferometer mirrors must be taken into account, and the appearance of optomechanical instabilities may jeopardize the stable operation of the next generation of interferometers. These instabilities are the result of a nonlinear coupling between the motion of the mirrors and the optical field, which modifies the effective dynamics of the mirror. Such 'optical spring' effects have already been demonstrated for the mechanical damping of an electromagnetic waveguide with a moving wall, the resonance frequency of a specially designed flexure oscillator, and the optomechanical instability of a silica microtoroidal resonator. Here we present an experiment where a micromechanical resonator is used as a mirror in a very high-finesse optical cavity, and its displacements are monitored with unprecedented sensitivity. By detuning the laser frequency with respect to the cavity resonance, we have observed a drastic cooling of the microresonator by intracavity radiation pressure, down to an effective temperature of 10 kelvin. For opposite detuning, efficient heating is observed, as well as a radiation-pressure-induced instability of the resonator. Further experimental progress and cryogenic operation may lead to the experimental observation of the quantum ground state of a micromechanical resonator, either by passive or active cooling techniques.

High-sensitivity optical monitoring of a micromechanical resonator with a quantum-limited optomechanical sensor.

We experimentally demonstrate the high-sensitivity optical monitoring of a micromechanical resonator and its cooling by active control. Coating a low-loss mirror upon the resonator, we have built an optomechanical sensor based on a very high-finesse cavity (30 000). We have measured the thermal noise of the resonator with a quantum-limited sensitivity at the 10(-19) m/sqrt[Hz] level, and cooled the resonator down to 5 K by a cold-damping technique. Applications of our setup range from quantum optics experiments to the experimental demonstration of the quantum ground state of a macroscopic mechanical resonator.

Spin squeezing and light entanglement in coherent population trapping.

We show that strong squeezing and entanglement can be generated at the output of a cavity containing atoms interacting with two fields in a coherent population trapping situation, on account of a nonlinear Faraday effect experienced by the fields close to a dark-state resonance in a cavity. Moreover, the cavity provides a feedback mechanism allowing to reduce the quantum fluctuations of the ground state spin, resulting in strong steady state spin squeezing.

Long-lived quantum memory with nuclear atomic spins.

We propose to store nonclassical states of light into the macroscopic collective nuclear spin (10(18) atoms) of a 3He vapor, using metastability exchange collisions. These collisions, commonly used to transfer orientation from the metastable state 2 3S1 to the ground state of 3He, can also transfer quantum correlations. This gives a possible experimental scheme to map a squeezed vacuum field state onto a nuclear spin state with very long storage times (hours).

Teleportation of an atomic ensemble quantum state.

We propose a protocol to achieve high fidelity quantum state teleportation of a macroscopic atomic ensemble using a pair of quantum-correlated atomic ensembles. We show how to prepare this pair of ensembles using quasiperfect quantum state transfer processes between light and atoms. Our protocol relies on optical joint measurements of the atomic ensemble states and magnetic feedback reconstruction.

Continuous variable entanglement using cold atoms.

We present an experimental demonstration of both quadrature and polarization entanglement generated via the interaction between a coherent linearly polarized field and cold atoms in a high finesse optical cavity. The nonlinear atom-field interaction produces two squeezed modes with orthogonal polarizations which are used to generate a pair of nonseparable beams, the entanglement of which is demonstrated by checking the inseparability criterion for continuous variables recently derived by Duan et al. [Phys. Rev. Lett. 84, 2722 (2000)]] and calculating the entanglement of formation [Phys. Rev. Lett. 91, 107901 (2003)]].

Polarization squeezing with cold atoms.

We study the interaction of a nearly resonant linearly polarized laser beam with a cloud of cold cesium atoms in a high finesse optical cavity. We show theoretically and experimentally that the cross-Kerr effect due to the saturation of the optical transition produces quadrature squeezing on both the mean field and the orthogonally polarized vacuum mode. An interpretation of this vacuum squeezing as polarization squeezing is given and a method for measuring quantum Stokes parameters for weak beams via a local oscillator is developed.

Experimental measurement of the dynamic photothermal effect in Fabry-Perot cavities for gravitational wave detectors.

We report the experimental observation of the frequency dependence of the photothermal effect. The measurements are performed by modulating the laser power absorbed by the mirrors of two high-finesse Fabry-Perot cavities. The results are very well described by a recently proposed theoretical model [M. Cerdonio, L. Conti, A. Heidmann, and M. Pinard, Phys. Rev. D 63, 082003 (2001)]], confirming the correctness of such calculations. Our observations and quantitative characterization of the dynamic photothermal effect demonstrate its critical importance for interferometric displacement measurements towards the quantum limit, as those necessary for gravitational wave detection.

A novel regulatory element in the dnmt1 gene that responds to co-activation by Rb and c-Jun.

Rb, c-Jun and dnmt1 play critical roles in the process of cellular differentiation. We demonstrate that a regulatory region of murine dnmt1 contains an element which is responsible for transactivation by Rb and c-Jun in P19 embryocarcinoma cells which is not observed in Y1 adrenocarcinoma cells. During differentiation of P19 cells, the induction of Rb and c-Jun coincides with an increase of dnmt1 mRNA. Using linker scanning mutagenesis we identify the element that is responsible for this activation to be a non-canonical AP-1 site. Our data is an example of how a proto-oncogene activates its downstream effectors by recruiting a tumor suppressor. This interaction of Rb and a proto-oncogene might play an important role in differentiation. The responsiveness of dnmt1 to this type of signal is consistent with an important role for regulated expression of dnmt1 during cellular differentiation.

Feedback regulation of DNA methyltransferase gene expression by methylation.

This paper tests the hypothesis that expression of the DNA methyltransferase, dnmt1, gene is regulated by a methylation-sensitive DNA element. Methylation of DNA is an attractive system for feedback regulation of DNA methyltransferase as the final product of the reaction, methylated DNA, can regulate gene expression in cis. We show that an AP-1-dependent regulatory element of dnmt1 is heavily methylated in most somatic tissues and in the mouse embryonal cell line, P19, and completely unmethylated in a mouse adrenal carcinoma cell line, Y1. dnmt1 is highly over expressed in Y1 relative to P19 cell lines. Global inhibition of DNA methylation in P19 cells by 5-azadeoxycytidine results in demethylation of the AP-1 regulatory region and induction of dnmt1 expression in P19cells, but not Y1 cells. We propose that this regulatory region of dnmt1 acts as a sensor of the DNA methylation capacity of the cell. These results provide an explanation for the documented coexistence of global hypomethylation and high levels of DNA methyltransferase activity in many cancer cells and for the carcinogenic effect of hypomethylating diets.

DNA methyltransferase is a downstream effector of cellular transformation triggered by simian virus 40 large T antigen.

This paper tests the hypothesis that DNA methyltransferase plays a causal role in cellular transformation induced by SV40 T antigen. We show that T antigen expression results in elevation of DNA methyltransferase (MeTase) mRNA, DNA MeTase protein levels, and global genomic DNA methylation. A T antigen mutant that has lost the ability to bind pRb does not induce DNA MeTase. This up-regulation of DNA MeTase by T antigen occurs mainly at the posttranscriptional level by altering mRNA stability. Inhibition of DNA MeTase by antisense oligonucleotide inhibitors results in inhibition of induction of cellular transformation by T antigen as determined by a transient transfection and soft agar assay. These results suggest that elevation of DNA MeTase is an essential component of the oncogenic program induced by T antigen.

Navigating ambulatory referral: a standardized process.

Significant funding and structure changes to healthcare in Ontario in the mid-90's led The Hospital for Sick Children in Toronto to examine patient referral processes. In an effort to streamline access and encourage more appropriate referrals, the hospital tested and implemented three major changes. This article outlines these changes using the PDSA (Plan, Do, Study, Act) improvement framework and summarizes the results from this project.