Distributed Water Pollution Source Localization with Mobile UV-Visible Spectrometer Probes in Wireless Sensor Networks

Pollution accidents that occur in surface water, especially in drinking water source areas, greatly threaten the urban water supply system. [...

The CXCR4 antagonist AMD3465 regulates oncogenic signaling and invasiveness in vitro and prevents breast cancer growth and metastasis in vivo.

CXCR4, the receptor for stromal-derived factor-1, is reportedly involved in breast carcinogenesis. However, the mechanisms through which CXCR4 contributes to breast cancer cell growth and metastases are poorly understood. In this study, we examined the putative in vitro and in vivo anti-cancer effects of the specific CXCR4 inhibitor AMD3465. Here, we report that AMD3465 triggers a reduction in breast cancer cell invasiveness in vitro, and promotes marked changes in oncogenic signaling proteins including a reduction in STAT3, JAK2, AKT, and CXCR4 phosphorylation and the reduced expression of GSK3 and cMYC. Using three breast cancer cell lines as murine syngeneic immunocompetent breast cancer models, we found that AMD3465 inhibited breast tumor formation and reduced tumor cell metastases to the lung and liver. Furthermore, treatment with AMD3465 significantly reduced the infiltration of myeloid CD11b positive cells at the aforementioned metastatic sites as well as the spleen implying this agent could regulate the formation of the tumor microenvironment and conceivably the premetastatic niche. In conclusion, our studies suggest that AMD3465 inhibits breast cancer growth and metastases by acting on tumor cells as well as immune cells that constitute the tumor microenvironment. This process appears to be regulated, at least in part, through the modulation of oncogenic signaling that includes the STAT3 pathway. Thus, CXCR4 could be a novel target for breast cancer therapy

AMD3465 Inhibits Breast Cancer Metastases.

<p>Site and number of metastases (Met.),</p>*<p>p<0.001 and **p<0.01 statically significant differences.</p

AMD3465 affects the <i>in vitro</i> invasiveness of 4T1 cells.

<p>4T1 cells in serum-free medium were seeded in matrigel transwells and allowed to migrate 48 h towards compartments with medium containing 10% FBS without (PBS) and with AMD3465 (2.5, 5, and 10 µM) present. All of the samples were conducted in triplicate and expressed as the mean value ± SD (error bars, **p<0.001).</p

AMD3465 inactivates CXCR4 in 4T1 tumors and slows tumor progression.

<p><b>A,</b> Tumor-bearing mice were injected with AMD3465 a single subcutaneous dose of 2.5 mg/kg. The tumor tissue was collected 1 h after treatment and sectioning was carried out. Immunohistochemical staining of pCXCR4, pAKT and pERK1/2 positive tumor cells can be seen in PBS controls compared to AMD3465 treated tumor sections. The slides were analyzed with an Olympus BX 41 microscope equipped with a digital camera (Olympus DP70). <b>B,</b> The top panel illustrates the BLI of 5 representative mice 1 d after injection of the 4T1 cells. The middle panel shows representative BLI in 5 mice treated with PBS 20 d after tumor injection, and the lower panel displays imaging of the 4T1 tumor masses following a similar exposure to AMD3465 (please see the Methods section for details). <b>C,</b> A bar graph representation of the end-point integrated photon (photons/cm²/sec) data collected in the experiment described in <b>B</b>. The tumor size was measured by BLI between control mice (n = 5) and AMD3465-treated mice (n = 10) and are expressed as the mean value ± SD (*p<0.05).</p

AMD3465 modulates intracellular oncogenic signaling mediators in mouse breast cancer cell lines.

<p><b>A,</b> A western blot analysis showing a 24-h knockdown of STAT3 expression using shRNA in the 4T1 cells and the concomitant abrogation of CXCR4 expression. <b>B,</b> Western blot analysis of oncogenic intermediates following a 24-h treatment of the 4T1 cells with 5 µM AMD3465. The band intensities ware normalized relative to β-actin expression and presented as % of Control in <b>C</b>. <b>D</b> and <b>E,</b> Western blot analyses of oncogenic intermediates following a 24-h treatment of the 4T1, 4T07, and 168Fran cells with 5 µM AMD3465.</p

AMD3465 reduces CD11b positive cells within metastatic lesions

<p>. <b>A,</b> A quantitative representations of CD11b positive cells in the lungs, liver, and spleen of immunocompetent syngeneic mouse model with the indicated cell line were calculated and the results are shown as bar graphs. The reductions in CD11b positive cells after a 14-d AMD3465 treatment in lung, spleen, and liver was observed in all three different breast cancer cell lines in an immunocompetent syngeneic mouse model. The percent positive cells were calculated based on total number of cells counted per image in triplicate samples, and expressed as the mean value ± SD (error bars) (**p<0.01 and ***P<0.001). <b>B,</b> A qualitative immunohistochemical depiction of the CD11b positive cells in representative lung tissues is shown for the 4T1, 4T07, and 168Farn cells in the immunocompetent syngeneic mouse model. <b>C,</b> Co-staining of metastatic nodules with CXCR4 (green fluorescence) and CD11b (red fluorescence) revealed ∼66% of the cells in the field are positive both for CD11b and CXCR4 (resulting yellow fluorescence) based on quantitative analysis of 10 images of the spleen tissue harvested from PBS-treated mice.</p

AMD3465 reduces tumor metastases in a syngenic breast cancer model.

<p>The size/number (indicated by arrows) of metastatic nodules in the 4T1 tumor bearing mice treated with PBS (control) or AMD3465 in both the lung and liver as determined by H&E staining. We also confirmed the metastatic nodules were GFP positive as were the primary tumors. A detailed treatment procedure for the metastasis assay is described in the Method section.</p