Metastatic properties of distinct phenotypic classes of lectin-resistant mutants isolated from murine MDAY-D2 cell line.

Single-step mutants were isolated from the murine metastatic MDAY-D2 cell line after selection in toxic concentrations of wheat-germ agglutinin. They were partially characterized by measuring their relative level of resistance to WGA, PHA, Con A, RIC, and LCA (Lec phenotype), and by comparing their karyotype and their ability to produce metastases upon transplantation into syngeneic DBA/2 mice. Based on their Lec phenotype, a total of 19 independent isolates were ranked into 10 distinct classes.

Characterization of in vitro immunoselected variants from a highly metastatic murine tumor for alterations in malignant behavior in vivo.

A new Ly-6.2- antigen-loss variant (called L61 -M1) of the highly metastatic DBA/2 mouse (Ly-6.2+) MDAY-D2 tumor has been obtained by means of a monoclonal anti-Ly-6.

Selection of strongly immunogenic "tum-" variants from tumors at high frequency using 5-azacytidine.

Highly immunogenic "tum-" (non-tumorigenic in normal syngeneic hosts) clonal variants can be selected from a variety of poorly immunogenic and highly tumorigenic mouse cell lines at very high frequencies (e.g., greater than 80%) after treatment in vitro with chemical mutagens such as ethyl methanesulfonate (EMS) or N-methyl-N'-nitro-N-nitrosoguanidine (MNNG).

Genotypic and phenotypic evolution of a murine tumor during its progression in vivo toward metastasis.

To follow the cellular progeny of the multiple-drug-marked benign murine tumor cell line MDW4 during its progression in vivo toward metastatic spread in DBA/2 mice, the following parameters were analyzed: retention of the drug-resistant markers ouabain resistance (OuaR) and thioguanine resistance (ThgR), lectin-resistance pattern (WGAR), and the karyotype of cell populations (and clones derived from these cells) removed at intervals from the solid tumor growing at the site of inoculation, as well as distant metastatic nodules. It was determined that the initially homogeneous inoculum composed of OuaR, ThgR, and WGAR hypotetraploid cells (mode: 68 +/- 2 chromosomes) was gradually overgrown and replaced by a new population of cells that were either OuaR or ouabain-sensitive but that became thioguanine-and lectin-sensitive and hyperploid (mode: 95 +/- 5). Regardless of the composition of the individual drug marker combinations, only cells with high chromosome contents were found to be able to disseminate to distant visceral organs and to rapidly produce metastases upon sc or iv reinjection.

Spontaneous fusion in vivo between normal host and tumor cells: possible contribution to tumor progression and metastasis studied with a lectin-resistant mutant tumor.

Previous studies demonstrated that growth in DBA/2 mice of MDW4, a wheat germ agglutinin-resistant (WGAr) mutant of the highly metastatic MDAY-D2 DBA/2 mouse tumor, led to the emergence of WGA-sensitive (WGAs) revertants having higher ploidy levels at the site of inoculation as well as at distant visceral metastases. The results implied that MDW4 was nonmetastatic but progressed to become metastatic in vivo only after a cellular change took place which was accompanied by extinction of the WGAr phenotype and acquisition of a higher number of chromosomes. Results presented here provide strong and direct evidence for the underlying mechanism being spontaneous cell fusion in vivo between the MDW4 (WGAr) tumor cells and normal host cells, at least some of which are of bone marrow origin.

Apparent reversion of stable in vitro genetic markers detected in tumour cells from spontaneous metastases.

The evolution towards more aggressive and autonomous behaviour of many cancerous tumours, often referred to as tumour progression, is thought to stem from the development of heterogeneity within the tumour cell population, combined with the continuous selection of progressively more malignant cellular phenotypes. During the course of the disease, the tumour cells show multiple phenotypic changes in a stepwise, but apparently random fashion, becoming more anaplastic, increasingly independent of growth controls and more metastatic. Several laboratories, including our own, have analysed aspects of tumour heterogeneity and cancer metastasis by selecting and studying the properties of lectin-resistant (LecR) membrane mutant tumour sublines; in a few cases, such variants have been claimed to be less tumorigenic or metastatic than the parental cells from which they were derived.

An examination of tumor antigen loss in spontaneous metastases.

Metastases arising from a subcutaneous injection of the DBA/2 tumor, MDAY-D2, as well as four drug-resistant variants (either wheat germ agglutinin-resistant, ouabain-resistant, or both, i.e., WGAR/OuaR) of MDAY-D2, were examined for the presence of a tumor-associated antigen (TAA).

Membrane-associated alterations detected in poorly tumorigenic lectin-resistant variant sublines of a highly malignant and metastatic murine tumor.

A number of wheat germ agglutinin-resistant (WGAR) variants of a highly malignant and metastatic mouse tumor (called MDAY-D2) were selected. Two of these, MDW1 and MDW3, were poorly tumorigenic in the normal DBA/2 host but grew well in highly immunosuppressed recipients. In contrast, MDAY-D2, MDW4, and MDW5 were all highly tumorigenic in both normal and immunosuppressed hosts.