Matrix metalloproteinases (MMPs) play an important role in tumor cell invasion and metastasis. These processes require the dissolution of the basement membrane and invasion of the stromal matrix (ECM), and are mediated by MMPs. Consequently, MMP inhibitors may be attractive as new anticancer agents. To examine the potential contribution of collagenase-1 (MMP-1) in invasion of stromal matrix, we used the highly invasive and metastatic breast cancer cell line MDA-MB-231 as a model system. These cells express procollagenase-1 constitutively and this expression can be repressed by all-trans retinoic acid. Invasion of these cells into a collagen type I matrix was assessed by scanning electron microscopy (SEM), and was quantitated with a computer program and confocal laser scanning microscopy (CLSM). We found that MDA-MB-231 cells freely invaded the collagen type I matrix, suggesting that these cells possess a mechanism for activating the latent collagenase-1. In contrast, down-regulation of collagenase-1 expression by all-trans retinoic acid caused these cells to become less invasive. To confirm a role for collagenase-1 in mediating collagen type I invasion, assays were carried out in the presence of FN-439, an inhibitor of collagenase-1 enzyme activity. We found that in the presence of the proteinase inhibitor, invasion of type I collagen by MDA-MB-231 cells was also reduced. These results indicate that collagenase-1 produced by the breast tumor cells may enhance stromal matrix degradation by enabling the tumor cells to modulate their own invasive behavior, and suggest that decreasing collagenase-1 levels may be effective in breast cancer therapy.
Generative cells from mature pollen grains of Haemanthus katherinae Baker (African blood lily) were isolated by means of a simple squash method and observed by differential interference contrast (DIC), fluorescence and polarizing microscopy. The isolated cells appeared structurally similar to those observed in vivo and gave no evidence of a typical cell wall. Their viability was confirmed using the fluorescein diacetate test. The cell shape changed rapidly as the sucrose concentration of the medium was varied. The squash method of isolating generative cells holds promise for the direct and experimental study of these cells, especially in the living state.
Generative cells were isolated from the pollen grains of three angiosperm species by a method similar to that previously reported for Haemanthus katherinae (Baker). Both the external appearance and the internal structure of the isolated generative cells were observed by light and scanning electron microscopy. The dynamic changes occurring in the cells after they had been liberated from the pollen grains were recorded by video-enhanced microscopy. The distribution of microtubules in the isolated cells was revealed by immunofluorescence.
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