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Rapid and quantitative in vitro measurement of cellular chemotaxis and invasion

An in vitro, fluorimetric method for cellular chemotaxis and invasion has been developed using a commercially available, disposable, 96-well chamber. This 4–18 hour microtiter chamber assay has a number of important advantages over existing methods. It does not require prior labeling of cells or radioactivity, and is rapid, automatable and quantitative. Cells are quantitated by a novel actin-based fluorescence tag as reported previously (Methods in Cell Science 17: 263–270, 1995). Following quantitation, cells are easily detectable by fluorescence microscopy. In addition, this assay conserves reagents due to its low volumes in the upper and lower chambers. The assay has been optimized using cultured human lung cancer cells to identify inhibitors or activators of directed cell migration. The effects of antibodies to agrVbeta3, agrVbeta5, and CD44 on the chemotaxis and invasion of A549 cultured lung tumor cells are reported.

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A Rapid in Vitro Assay for Quantitating the Invasive Potential of Tumor Cells

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 lOT’/i 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.

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Note: do not autoclave acrylic chambers as this article recommends.

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Insulin-like Growth Factor (IGF)-binding Protein-4 Inhibits Colony Formation of Colorectal Cancer Cells by IGF-independent Mechanisms

Effects of insulin-like growth factor-binding protein-4 (IGFBP-4) on proliferation, colony formation, and cell migration were assessed in IGF-sensitive and -insensitive colorectal cancer cell lines. In IGF-insensitive Isreco-1 cells, overexpression of IGFBP-4 reduced colony formation but not cell proliferation and migration, whereas exogenous IGF-II had no effect. In IGF-dependent LS1034 cells, IGFBP-4 inhibited all parameters of growth tested, whereas IGF-II partially restored reduced proliferation and cell migration only. In Isreco-2 cells, which lack endogenous IGF expression but are IGF sensitive, colony formation was also reduced by IGFBP-4. Therefore, specific parameters of malignant progression of colon carcinoma cells are distinctly affected by IGF-dependent and IGF-independent effects of IGFBP-4.