Phosphoinositide 3-kinase signaling pathway mediated by p110α regulates invadopodia formation

Inhibition of p110α or of the downstream PI3K signaling pathway components PDK1 and Akt, as well as phosphoinositide sequestration, blocks invadopodia formation in breast cancer cells

Thyrotoxic and pheochromocytoma multisystem crisis: a case report

Abstract Background Thyrotoxic crisis and pheochromocytoma multisystem crisis are rare, life-threatening, emergency endocrine diseases with various clinical manifestations. Here we report a case of a patient who simultaneously developed thyrotoxic crisis and pheochromocytoma multisystem crisis and required intensive cardiovascular management. Case presentation A 60-year-old Asian man experienced nausea and vomiting, and subsequently developed dyspnea and cold sweats while farming. His serum free thyroxine, free triiodothyronine, and thyrotropin receptor antibody levels were elevated at 2.9 ng/dL, 7.2 pg/dL, and 4.7 IU/L, respectively. Serum thyrotropin levels were suppressed at less than 0.01 μIU/mL. Thyroid echography demonstrated no thyroid swelling (23 × 43 mm). A whole body computed tomography was performed for systemic evaluation. This revealed exophthalmos and a mass of size 57 × 64 mm in the anterior pararenal space. Based on these findings, we made an initial diagnosis of thyrotoxic crisis secondary to exacerbation of Grave’s hyperthyroidism. Treatment was begun with an iodine agent at a dose of 36 mg/day, thiamazole at a dose of 30 mg/day, and hydrocortisone at a dose of 300 mg daily for 3 consecutive days. To control tachycardia, continuous intravenously administered propranolol and diltiazem infusions were given. At the same time, small doses of doxazosin and carvedilol were used for both alpha and beta adrenergic blockade. On hospital day 5, his blood pressure and serum catecholamine concentrations (adrenalin 42,365 pg/mL, dopamine 6409 pg/mL, noradrenalin 72,212 pg/mL) were still high despite higher beta blocker and calcium channel blocker doses. These findings contributed to the diagnosis of pheochromocytoma multisystem crisis with simultaneous thyrotoxic crisis. We increased the doses of doxazosin and carvedilol, which stabilized his hemodynamic status. On hospital day 16, metaiodobenzylguanidine scintigraphy showed high accumulation in the right adrenal gland tumor. After retroperitoneal laparoscopic adrenalectomy on hospital day 33, his condition stabilized. He was discharged on hospital day 58. Conclusions Since he required more intensive cardiovascular management for thyrotoxic crisis, beta blockade was increased under intensive care unit monitoring even though initial alpha blockade is recommended in pheochromocytoma. When these crises occur simultaneously, cardiovascular management can be very challenging

Dasatinib inhibits cell aggregation, migration, and invasion in vitro.

<p>A, Fluorescent images of individually cultured and cocultured 44As3 and CaF37 cells on 3D Matrigel in the presence or absence of dasatinib (1 or 10 µM) for 2 days. B, Quantification of the areas and number of cell clusters. Bars show mean ± SEM (<i>n</i> = 48–502 for the cluster area, 6 for the number of clusters). *, <i>p</i><0.05; **, <i>p</i><0.001 by Student's <i>t</i>-test. C, Total and net migration distances of cells per hour were measured from time-lapse movies. Bars show mean ± SEM (<i>n</i> = 40). *, <i>p</i><0.0001 by Student's <i>t</i>-test. D, Invasion depth of the invasive foci or cell clusters. Bars show mean ± SD (<i>n</i> = 20). *, <i>p</i><0.0001 by Student's <i>t</i>-test.</p

Dasatinib suppresses peritoneal dissemination of SGC cells and their association with stromal fibroblasts in vivo.

<p>A, 44As3 cells were intraperitoneally injected into nude mice and DMSO or dasatinib was administered via intraperitoneal injection. The number of mesentery nodules was calculated as described in the Materials and Methods. Bars show mean ± SEM (<i>n</i> = 10). *, <i>p</i><0.005 by Mann-Whitney test. B, Representative macroscopic views of metastatic tumor nodules (arrowheads) formed in the mesentery. C, Immunofluorescence analysis of the mouse mesenteries bearing tdTomato-labeled 44As3 tumor nodules. Arrowheads denote the regions where FSP1 positive stromal fibroblasts were accumulated around tumor nodules. D, Mesentery nodules were stained with hematoxylin and eosin and anti-αSMA antibody for histological examination.</p

Rock and Src regulate the formation of invasive foci.

<p>A, 44As3 and CaF37 cells were plated onto 3D Matrigel in the presence or absence of H1152 (10 µM) or dasatinib (10 µM) for 2 days. B, Quantification of the number of invasive foci. Bars show mean ± SEM (<i>n</i> = 3). *, <i>p</i><0.05 by Student's <i>t</i>-test. C, Dose response effects of H1152 and dasatinib on cell growth of 44As3 and CaF37 cells and the formation of invasive foci. Bars show mean ± SD (<i>n</i> = 8 for cell growth and 3 for invasive foci). *, <i>p</i><0.05 by Student's <i>t</i>-test. D, Immunoblot analysis of 44As3 and CaF37 cells that were cocultured and treated with H1152 or dasatinib. E, The effect of H1152 and dasatinib on remodeling of the gelatin matrix. F, Quantification of the areas of gelatin disruption. Bars show mean ± SEM (<i>n</i> = 3). *, <i>p</i><0.0005 by Student's <i>t</i>-test.</p

Stromal Fibroblasts Mediate Extracellular Matrix Remodeling and Invasion of Scirrhous Gastric Carcinoma Cells

<div><p>Scirrhous gastric carcinoma (SGC) has the worst prognosis of all gastric cancers, owing to its rapid expansion by invasion and frequent peritoneal dissemination. Due to the increased proliferation of stromal fibroblasts (SFs) that occurs within SGC lesions and the peritoneal metastatic sites, SFs have been proposed to support the progression of this disease. However, the biological and molecular basis and the pathological role of the intercellular interaction between SGC cells and SFs remain largely unknown. In this study, we investigated the role of SFs in the invasion of the extracellular matrix (ECM) by SGC cells. When SGC cells were cocultured with SFs derived from SGC tissue on three-dimensional (3D) Matrigel, they were attracted together to form large cellular aggregates that invaded within the Matrigel. Time-lapse imaging revealed that this process was associated with extensive contraction and remodeling of the ECM. Immunofluorescence and biochemical analysis showed that SGC cells stimulate phosphorylation of myosin light chain and actomyosin-mediated mechanical remodeling of the ECM by SFs. By utilizing this assay system for inhibitor library screening, we have identified several inhibitors that potently suppress the cooperation between SGC cells and SFs to form the invasive structures. Among them, a Src inhibitor dasatinib impaired the interaction between SGC cells and SFs both in vitro and in vivo and effectively blocked peritoneal dissemination of SGC cells. These results indicate that SFs mediate mechanical remodeling of the ECM by SGC cells, thereby promoting invasion and peritoneal dissemination of SGC.</p></div

Effect of dasatinib on peritoneal dissemination of 44As3 cells.

<p>Number of mice bearing ascites or tumor at the indicated site per total number of mice.</p