Quantitative analysis of cancer invasion in vitro: comparison of two new assays and of tumour sublines with different metastatic capacity

Abstract

Various murine tumour sublines which differed considerably in their in vivo metastatic capacity were tested in vitro for their ability to invade normal tissue. For this purpose we developed two quantitative tests, a Boyden chamber endothelial cell invasion assay and a brain tissue microsphere invasion assay. The invasion of [75Se]methionine-prelabelled tumour cells into the normal tissues was followed by measuring the percentage of tumour-associated label in the brain microspheres or the endothelial monolayers after 12-48 h of co-cultivation. Clear and comparable differences existed in both assays between the amount of radiolabel found in the normal tissues after a co-cultivation with the different tumour lines. In three of the four tumour lines invasiveness correlated with metastatic capacity. The fourth line, a plastic adherent variant, was highly invasive but low metastatic. The ability of tumour cells to invade normal tissue, therefore, while necessary for the generation of metastases, is not in itself sufficient. Since both assays are independent of time-consuming histological sectioning and staining and allow a quantitative determination of invasive capacity of tumour cells grown as single cell suspensions they appear well suited for experimental manipulation and for screening of anti-invasive drugs.

Organ specificity of tumor metastasis: role of preferential adhesion, invasion and growth of malignant cells at specific secondary sites

“…In general, only the highly metastatic cells were capable of attaching to and invading the capillary surfaces. Although brain tissue is difficult to culture for tumor cell adhesion and invasion studies, Waller et al [253] mechanically dispersed brain through 200/zm nylon mesh to form tissue microspheres that could be maintained in tissue culture for several days, and Wang and Nicolson [254] cultured small pieces of brain tissue on cellulose polyacetate strips for tumor cell adhesion and invasion studies. Other tissues, such as syngeneic bladder, have been used to select invasive variants [96,97], or to monitor the invasive behaviors of bladder carcinoma cells [255].…”

Origin and biology of cancer metastasis

“…Instead, passive mechanical processes such as turbulence probably are responsible for a high rate of tumor cell death. Unlike blood cells, tumor cells are not sufficiently deformable to survive the high shear forces imposed on them by the microcirculation (81,93,98). The few cells that do survive circulatory transport may do so because they possess enhanced deformability (81,981 or form an embolus.…”

“…The cells are then tested in the appropriate host to determine their metastatic potential. This method has been used to examine whether properties as diverse as resistance to T lymphocytes (16,19,28,42,58,93), adhesive characteristics (a), invasive capacity (34,66,69,73), resistance to various lectins (42,43,79), and resistance to natural killer cells (33) are important in metastasis.…”

“…The cells are then tested in the appropriate host to determine their metastatic potential. This method has been used to examine whether properties as diverse as resistance to T lymphocytes (16,19,28,42,58,93), adhesive characteristics (a), invasive capacity (34,66,69,73), resistance to various lectins (42,43,79), and resistance to natural killer cells (33) are important in metastasis.One obvious criticism of these studies is that the surviving tumor cells may have arisen as a result of adaptive rather than selective processes. The first experimental proof for metastatic heterogeneity in neoplasms was provided by Fidler and Kripke in 1977 in work with the murine B16 melanoma (18).…”

Determination of cell-free interleukin 2 receptor level in the serum of normal animals and of animals bearing IL-2 receptor positive tumours with high or low metastatic capacity

“…The spontaneous high metastatic variant ESb is most likely derived from a spontaneous fusion of the parental Tlymphoma Eb with a host macrophage (Larizza et al, 1984). The two lines were found to differ in many cell surface properties including expression of differentiation antigens , of composition of membrane glycoproteins (Schwartz et al, 1984), glycolipids (Murayama et al, 1986) and in the shedding of membrane vesicles (Barz et al, 1985;Schirrmacher & Barz, 1986 (Waller et al, 1986). In vivo, however, the ESb-M line showed greatly decreased overall malignancy (Benke et al, 1987).…”