Unlinked regulation of the sensitivity of primary glucocorticoid-inducible responses in mouse mammary tumor virus infected Fu5-5 rat hepatoma cells.

The enzyme tyrosine aminotransferase (TAT) is induced by unusually low concentrations of glucocorticoids in Fu5-5 cells. We have isolated clones of Fu5-5 cells infected with mouse mammary tumor virus (MMTV) in order to simultaneously compare the glucocorticoid regulation of the host cell gene, TAT, with that of another primary inducible gene, MMTV. In the two clones that were examined in detail, MMTV RNA induction occurred at 4- to 11-fold higher concentrations of dexamethasone than those needed for induction of TAT mRNA. Furthermore, the amount of agonist activity displayed by the irreversible antiglucocorticoid dexamethasone 21-mesylate was greater for the induction of TAT mRNA than for MMTV RNA. These results extend our previous observations of unequal sensitivity of induction of TAT enzyme activity in two hepatoma cell lines and show that differential glucocorticoid regulation of gene induction within the same cell can occur at a pretranslational step. The present data also indicate that the unusual properties of TAT gene induction are not shared by all primary, glucocorticoid-inducible responses of the same cell and imply that additional factors mediating differential regulation of glucocorticoid-responsive genes are involved.

Heterogeneity of in vitro growth pattern of megakaryocyte progenitors (CFU-M) in myeloproliferative disorders.

In groups of 26 patients with myeloproliferative disorders (MPD), 8 with chronic myelogenous leukaemia (CML); 8 with polycythaemia vera (PV); 10 with essential thrombocythaemia (ET); and 6 patients with reactive thrombocytosis (RT), we studied the growth characteristics of bone marrow CFU-M in agar culture. The bone marrows from all the patients with MPD formed so called endogenous CFU-M colonies, in the absence of PHA-LCM, that increased in a dose-dependent manner with the addition of increasing concentrations of normal human AB-citrated plasma (NH-ABCP), while the bone marrows from all the patients with RT and from healthy controls formed few or no endogenous CFU-M colonies. In MPD, the endogenous CFU-M growth was enhanced by normal T cells in a dose-dependent fashion, and was decreased with the depletion of T cells from the marrow cells. These results suggest that the formation of endogenous CFU-M colonies is caused by hypersensitivity of CFU-M in MPD to NH-ABCP, which may contain a small amount of Meg-CSF, and/or by in vitro T cell stimulation. Among MPD, the endogenous CFU-M growth in ET was significantly lower than that of other MPD patients; however, the total number of ET CFU-M grown in the presence of PHA-LCM was the highest. These data show that the bone marrow CFU-M in MPD are heterogeneous with respect to in vitro growth pattern or sensitivity to exogenous Meg-CSF.

[Clonal structure of tumors: difference between clones based on the ability to secrete growth-regulating factors and to react to them]. / Klonal'naia struktura opukholi: razlichiia mezhdu klonami po sposobnosti vydeliat' faktory, reguliruiushchie razmnozhenie, i reagirovat' na nikh.

Conditioned medium isolated from different clones of tumour cell culture PS-103 (sarcoma induced in CBA mouse by plastic film implantation) varied in the ability to stimulate cell proliferation in semi-solid medium. The clones differed also in their response to the medium conditioned by culture PS-103: some clones responded by increased proliferation in semi-solid medium, some were inhibited and some failed to respond.

Effect of nerve growth factor producing cells on anaplastic glioma and pheochromocytoma clones: involvement of other factors.

When the NGF-secreting C-6 rat glioma cells were intracerebrally injected with F98 rat anaplastic glioma cells into rats syngeneic for the F98 cells, an increased mean survival time was observed for rats developing tumors compared with those injected with only anaplastic glioma cells. Thirty percent of the rats injected with both cell types showed no signs of tumor at 90 days. Pretreatment of the anaplastic glioma cells with conditioned medium of C-6 cells did not duplicate these results unless the C-6 cells were pretreated with 17 beta-estradiol, which appears to induce secretion of an adhesion factor as well as NGF. These rats survived even longer and 33% were free of tumors at 90 days. Histological examination of tumors of the nonsurviving rats revealed that they were basically well differentiated with only small anaplastic areas remaining. Both NGF and conditioned medium from C-6 and another NGF-secreting line, S-180 mouse sarcoma, induce process formation in F98 cells and in PC12 pheochromocytoma cells, but the processes that appear following NGF exposure are morphologically different from those induced by conditioned medium. Conditioned -medium-treated cells also have a flatter appearance. In F98 cells, NGF takes longer to induce processes than does conditioned medium. The NGF-induced effects observed in both cells are neutralized by anti-NGF IgG, but those induced by conditioned media are not. Nerve growth factor (NGF) induced increased adhesiveness in F98 rat anaplastic glioma and PC12 rat pheochromocytoma cells. Conditioned media are not. Nerve growth factor induced increased adhesive-and PCuced effects observed in both cells are neutralized by anti-NGF IgG, but those induced by conditioned media are not. Nerve growth factor (NGF) induced increased adhesiveness in F98 rat anaplastic glioma and PC12 rat pheochromocytoma cells. Conditioned media are not. Nerve growth factor induced increased adhesive-and PCuced effects observed in both cells are neutralized by anti-NGF IgG, but those induced by conditioned media are not. Nerve growth factor (NGF) induced increased adhesiveness in F98 rat anaplastic glioma and PC12 rat pheochromocytoma cells. Conditioned media are not. Nerve growth factor induced increased adhesive-and PC12 cells. Anti-NGF IgG did not influence this effect on F98 cells and only partially neutralized the effect on PC12 cells, indicating that other factors may be operative in this system. Conditioned medium collected from C-6 cells pretreated with 17 beta-estradiol induced the highest degree of adhesiveness observed in both F98 and PC12 cells, and this action was unaffected by anti-NGF IgG in either case. Conditioned media from other cell lines, a variety of selected proteins, and dBcAMP did not induce increased adhesiveness. The factor responsible for this effect is nondialyzable, heat-sensitive, and ammonium-sulfate-precipitable, and its secretion appears to be stimulated by 17 beta-estradiol.

Genetic analysis of tumorigenesis: I. Expression of tumor-forming ability in hamster hybrid cell lines.

Two new fibroblasts cell lines from a male Chinese hamster embryo are described: one tumorigenic in nude mice (CHEF 16-2) and the other not (CHEF 18-1). Both lines have a stable diploid mode and mean of 22 with 10 pairs of homologous autosomes. The cell lines differ unambiguously in cloning ability in methylcellulose, colony morphology, and tumorigenicity; the expression of these traits was examined in a set of 18-1 X 16-2 hybrid clones. The results show initial suppression of tumorigenicity and anchorage independence, followed by growth in the nude mouse of subsets of cells with chromosomes reduced to the diploid range. Further studies are in progress to establish whether ability to grow in methylcellulose and in the nude mouse segregate coordinately at the cellular level, and whether this segregation has an identifiable chromosomal basis.