Abstract
Three human malignant melanoma cell strains (C8146A, C8146C, and C83-2CY), three established human melanoma cell lines (A375P, A375M, and C8161), and one selected human melanoma subline (A375P-5) were studied to determine if invasion through a reconstituted basement membrane-coated filter (RBMF), which selects the most aggressively invasive cells, would also modulate the cloning efficiency of these cells in soft agar. With the use of the Membrane Invasion Culture System (MICS), all cell strains tested showed a significant increase in cloning efficiency (1.05–9.3-fold) following transit through the RBMF when compared to unmanipulated populations. The established cell lines (A375P, A375M, and C8161) and the A375P-5 subline showed either a decrease or unaltered status in cloning efficiency after invasion. However, all cells demonstrated a consistent decrease in clonogenicity following transit through an uncoated filter compared with RBMFs, thus suggesting the influence of the extracellular matrix on tumor cell clonogenic properties. In general, the established cell lines were more clonogenic before invasion of the RBMF compared with the cell strains, and no correlation was found between clonogenic potential and invasive or metastatic capability. These data may provide important insight into the underlying mechanisms of tumor cell invasion and the subsequent formation and dissemination of metastases in vivo.
Highlights
The most important clinical problem of cancer is metastasis - the migration of tumor cells through the vasculature and tissue parenchyma that gives rise to additional tumors in the body
An additional subline was used in this study - A375P5, which was derived by allowing A375P cells to interact with a reconstituted basement membrane-coated filter (RBMF) in MegaMICS chambers for 24 h, collecting the highly invasive subpopulation that invaded the RBMF, and subsequently reintroducing the subpopulation to additional RBMFs and repeating the sequence a total of five times
C8146A cells collected from the undersurface of RBMFs showed a 1.5% increase in cloning efficiency compared with pre-invasive cells grown on plastic, a 0.9 % increase compared with pre-invasive cells grown on RBM-coated culture dishes, and a 7.8% increase compared with pre-invasive cells attached to the uppersurface of the RBMFs during the 48 h MegaMICS assay
Summary
The most important clinical problem of cancer is metastasis - the migration of tumor cells through the vasculature and tissue parenchyma that gives rise to additional tumors in the body. Of the many types of cancers, malignant melanoma, considered one of the most severe and painful neoplastic diseases, is increasing in incidence across the globe. 0 1989 Elsevier Scientific Publishers Ireland Ltd. Published and Printed in Ireland are forced to employ relatively nonspecific therapeutic regimes in the treatment of this malignant disease. The acquisition of the capacity to invade or penetrate a basement membrane (BM) is of primary importance in both the development of malignant melanoma and the genesis of many kinds of advanced malignancy because the BM is the most ubiquitous biological barrier that a wandering tumor cell is likely to encounter during intravasation, extravasation and dissemination in the human body [ 13,151. Within a primary tumor are subpopulations of cells [7] containing some cells capable of invasion, other cells capable of invasion and metastasis [3], and still other cells with unknown capabilities
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