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  • Inhibition of adhesion, invasion, and metastasis by antibodies targeting CEACAM6 (NCA-90) and CEACAM5 (Carcinoembryonic Antigen). 16204051

    CEACAM5 and CEACAM6 are overexpressed in many cancers and are associated with adhesion and invasion. The effects of three monoclonal antibodies targeting different epitopes on these antigens (NH2-terminal [MN-3] and A1B1 domains [MN-15] shared by CEACAM5 and CEACAM6 and the A3B3 domain [MN-14] restricted to CEACAM5) were evaluated in migration, invasion, and adhesion assays in vitro using a panel of human pancreatic, breast, and colonic cancer cell lines, and in the GW-39 human colonic micrometastasis model in vivo. MN-3 Fab' and MN-15 Fab' were both effective at inhibiting cell migration. MN-15 Fab' treatment inhibited invasion, reducing cell penetration through an extracellular matrix (ECM). MN-3 Fab' also decreased invasion but was less effective than MN-15 Fab' in four of five cell lines. All three monoclonal antibody (mAb) Fabs decreased adhesion of tumor cells to endothelial cells by 49% to 58%. MN-15 Fab' but not MN-3 or MN-14 Fabs induced a decrease in adhesion of three of six cell lines to the ECM protein, fibronectin, but adhesion to vitronectin, laminin, collagen-I, and collagen-IV was not affected. In vivo studies showed that treatment with MN-3 Fab' or MN-15 Fab' of mice implanted with GW-39 human colonic cancer cells increased their survival (P < 0.025 and P < 0.01, respectively). These studies show that antibody Fabs that target either CEACAM5 or CEACAM6 affect cell migration, cell invasion, and cell adhesion in vitro, and that MN-15 and MN-3 Fabs have antimetastatic effects in vivo, resulting in improved survival of mice with metastases. Thus, blocking the N and A1B1 domains of CEACAM5/CEACAM6 can impede the metastatic process.
    Document Type:
    Reference
    Product Catalog Number:
    ECM551
    Product Catalog Name:
    QCM™ Collagen Cell Invasion Assay, 24-well (8 µm), Colorimetric

  • Plumbagin, a plant derived natural agent inhibits the growth of pancreatic cancer cells in in vitro and in vivo via targeting EGFR, Stat3 and NF-κB signaling pathways. 22322442

    Pancreatic cancer (PC) is the most aggressive malignant disease, ranks as the fourth most leading cause of cancer-related death among men and women in the United States. We present here that plumbagin (PL), a quinoid constituent isolated from the roots of the medicinal plant Plumbago zeylanica L, inhibits the growth of PC cells both in vitro and in vivo model systems. PL treatment induces apoptosis and inhibits cell viability of PC cells (PANC1, BxPC3 and ASPC1). In addition, i.p. administration of PL (2 mg/kg body weight, 5 days a week) in severe combined immunodeficiency (SCID) mice beginning 3 days after ectopic implantation of PANC1 cells resulted in a significant (P < 0.01) inhibition of both tumor weight and volume. PL treatment inhibited (1) constitutive expression of epidermal growth factor receptor (EGFR), pStat3Tyr705 and pStat3Ser727, (2) DNA binding of Stat3 and (3) physical interaction of EGFR with Stat3, in both cultured PANC1 cells and their xenograft tumors. PL treatment also inhibited phosphorylation and DNA-binding activity of NF-κB in both cultured PC cells (PANC1 and ASPC1) and in PANC1 cells xenograft tumors. Downstream target genes (cyclin D1, MMP9 and Survivin) of Stat3 and NF-κB were similarly inhibited. These results suggest that PL may be used as a novel therapeutic agent against human PC. Published 2012 Wiley-Liss, Inc. This article is a US Government work, and, as such, is in the public domain in the United States of America.
    Document Type:
    Reference
    Product Catalog Number:
    ECM551
    Product Catalog Name:
    QCM™ Collagen Cell Invasion Assay, 24-well (8 µm), Colorimetric

  • A rapid in vitro assay for quantitating the invasive potential of tumor cells. 2438036

    We have reconstituted a matrix of basement membrane onto a filter in a Boyden chamber and assessed the ability of various malignant and nonmalignant cells to penetrate through the coated filter. Cells from all the malignant cell lines tested were able to cross the matrix in 5-6 h, whereas human fibroblasts as well as mouse 3T3 and 10T1/2 cell lines, which are not tumorigenic, were not invasive. In addition, normal primary prostate epithelial cells and benign prostatic hyperplasia cells were not invasive when tested in this assay, whereas malignant prostate carcinoma cells were highly invasive. Parallel experiments with these prostatic cells using the intrasplenic assay for metastasis detection in the nude mouse confirmed the benign behavior of the former cells and the metastatic phenotype of the latter ones. These results suggest that this in vitro test allows the rapid and quantitative assessment of invasiveness and a means to screen for drugs which alter the invasive phenotype of tumor cells.
    Document Type:
    Reference
    Product Catalog Number:
    ECM551
    Product Catalog Name:
    QCM™ Collagen Cell Invasion Assay, 24-well (8 µm), Colorimetric

  • MiR-145 inhibits tumor angiogenesis and growth by N-RAS and VEGF. 22592534

    MiR-145 is known as a tumor suppressor in numerous human cancers. However, its role in tumor angiogenesis remains poorly defined. In this study, we found that miR-145 was significantly downregulated in breast cancer tissues by using 106 cases of normal and cancer tissues as well as in breast cancer cells. MiR-145 exhibited inhibitory role in tumor angiogenesis, cell growth and invasion and tumor growth through the post-transcriptional regulation of the novel targets N-RAS and VEGF-A. In addition, we provide evidence that the expression levels of miR-145 correlate inversely with malignancy stages of breast tumors, although there is no association between miR-145 levels and hormone receptor levels in breast cancer. Taken together, these results demonstrate that miR-145 plays important inhibitory role in breast cancer malignancy by targeting N-RAS and VEGF-A, which may be potential therapeutic and diagnostic targets.
    Document Type:
    Reference
    Product Catalog Number:
    ECM551
    Product Catalog Name:
    QCM™ Collagen Cell Invasion Assay, 24-well (8 µm), Colorimetric

  • A new in vitro assay for quantitating tumor cell invasion. 2722434

    The attachment to and penetration of basement membranes by tumor cells is required to complete the metastatic cascade which culminates in the establishment of secondary tumor foci. Therefore, basement membranes are critical barriers to the passage of disseminating tumor cells. We have developed a simple, in vitro model using matrigel-coated transwell chambers (Costar) for use in a tumor cell invasion assay. Two variants of the K1735 UV-induced murine melanoma cell line were assayed for their invasive capabilities and compared with their ability to colonize the lung in an experimental metastasis assay. The K1735-M2 cells, which are highly metastatic in vivo, invaded through basement membrane matrigel at a significantly higher rate than the low metastatic cells, K1735-16, in a 72-hour assay. As a negative control, normal murine fibroblasts were incapable of penetrating the barrier. Tumor cell invasion in vitro correlated with lung colonization in vivo. Therefore, this model may provide a valuable tool to study the mechanisms involved in the pathogenesis of tumor cell invasion during hematogenous dissemination.
    Document Type:
    Reference
    Product Catalog Number:
    ECM551
    Product Catalog Name:
    QCM™ Collagen Cell Invasion Assay, 24-well (8 µm), Colorimetric

  • User Manual-ECM551

    Document Type:
    User Guide
    Product Catalog Number:
    ECM551
    Product Catalog Name:
    QCM™ Collagen Cell Invasion Assay, 24-well (8 µm), Colorimetric

  • Use of a reconstituted basement membrane to measure cell invasiveness and select for highly invasive tumor cells. 3455782

    Malignant cells must traverse basement membranes during their migration to sites distant from the primary tumor. Since basement membranes are thought to be a critical barrier to the passage of tumor cells, we have constructed a model basement membrane-stromal matrix consisting of laminin and type IV collagen reconstituted onto a disk of type I collagen for use in an in vitro assay of invasiveness. Metastatic tumor cells and leukocytes are able to cross this barrier, whereas nonmetastatic tumor cells, fibroblasts, and epidermal cells cannot penetrate it. Those tumor cells that penetrate the barriers were found, when isolated and subcultured, to be more invasive and to produce more metastases than the parental population. This assay system should be useful for studying the invasiveness of tumor cells and for isolating highly invasive variants.
    Document Type:
    Reference
    Product Catalog Number:
    ECM551
    Product Catalog Name:
    QCM™ Collagen Cell Invasion Assay, 24-well (8 µm), Colorimetric