Jadwiga Koziorowska

Studies on adaptation to adriamycin in cells pretreated with hydrogen peroxide

Various investigations have reported the occurrence in bacterial and mammalian cells of an adaptive response to the toxic effects of oxidants or agents that cause oxidation via redox reactions. In our previous study, it was shown that several cell lines pretreated with a low dose of hydrogen peroxide (H2O2) exhibited an adaptive response to subsequent high doses of adriamycin (ADR), whereas other cell lines did not. Based on the observation that the cell lines utilized differed in their sensitivity towards adriamycin, we undertook the present investigation with the goal of evaluating possible relationships between the levels of antioxidant enzymes and sensitivity towards adriamycin. Another aim was to determine relationships between the inducibility of these enzymes and the occurrence of adaptation. We utilized African Green monkey kidney (V3), human embryo (CLV98), human melanoma (ME18), and Chinese hamster ovary (CHO) cell lines and experimentally developed adriamycin-resistant human melanoma (ME18/RN) and Chinese hamster ovary (CHO/RN) cell sublines. Cytotoxicity was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and trypan blue exclusion. The levels of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were determined in the same kind of experiment as that revealing the occurrence of adaptation. The rank order established for catalase activities was similar to that for sensitivity towards adriamycin. Aberrant increases in the tested enzymes were demonstrated in experimental groups of all kinds of cells. We conclude that in our cell systems catalase is a major determinant of adriamycin resistance. Whether the occurrence of the adaptive response under study is dependent on the contribution of catalase, itself dependent on the degree of resistance to the drug, is discussed.

Influence of retinoic acid on protein synthesis and transport of l-methionine in cultured L cells

The effects of retinoic acid (RA) on cell proliferation, activity of acid phosphatase, protein synthesis and methionine uptake were studied in transformed murine LPA cells. Early inhibition of protein synthesis was demonstrated under experimental conditions in which the rate of cell proliferation was diminished and non-specific effects of vitamin action could be excluded. Measurements of L-methionine uptake revealed a decrease to approximately one-half of that in control cultures after treatment with RA at the concentrations of 5 X 10(-5) M and 10(-4) M.

Vaccinia virus in organ cultures of an adult rabbit's skin

Abstract Explants of full-thickness skin of adult rabbits were infected with high and low doses of vaccinia virus and cultivated as organ cultures. The progressive cytological alterations were confined to epithelial cells and resembled those described by several authors in rabbits in vivo. Thus, organ cultures from differentiated tissues provide a good model for investigations of hostvirus relationships. Histological examination and counts of mitotic cells have shown that infection in vitro with vaccinia results in stimulation of epidermal growth and brings about intra-epidermal necrosis. A similar duality of effects on epidermal cells is manifested with vaccinia virus in vivo. Infected cultures were compared with control cultures as to their mitotic potential and type of keratinization. Marked differences were found indicating that infection with vaccinia affects growth and differentiation of the epidermis in a way which is dependent upon the amount of virus particles introduced initially into the culture medium.

Adaptive response towards adriamycin in vitro: circumvention with verapamil.

In an attempt to identify mechanisms of adaptive response to adriamycin (ADR), we have earlier isolated ADR-resistant cell lines CHO/R and ME18/R by short-term pulse exposures of parent cell lines to this drug, followed by single-cell cloning. The results presented in this study have shown that the development of resistance to ADR was accompanied by cross-resistance to vinblastine and methotrexate. The resistance of tested cell lines towards ADR was substantially reversed by verapamil (VPL) at non-toxic concentrations. VPL abolished also the capability of these cell lines to express adaptive response after treatment of the cells with a conditioning dose of ADR. From the results of our study, we conclude that similar characteristics play a role in the mechanism of the phenomenon of adaptive response as in the mechanism of pleiotropic multidrug resistance.

Methionine dependence of virus-infected cells

Mouse embryo cells nonproductively infected with human cytomegalovirus differed from noninfected cells by the impaired ability to grow in the medium containing homocysteine instead of methionine. Virus infection of mouse embryo cells grown in both kinds of media resulted in the increase of protein synthesis. In the infected cells grown on homocysteine this increase was followed by a quick decrease. The effects of homocysteine substitution could be abolished by the addition of low amounts of methionine (0.1 mM). Methionine uptake in the infected cells grown on homocysteine for 48 h was significantly higher than that in the noninfected cells.

Study on the interaction of ions of transient metals with ascorbic acid in the presence of different scavengers of active oxygen species in SOS chromotest.

SOS chromotest was employed to study the interaction of ascorbic acid with free ions of transient metals in the presence of added catalase, superoxide dismutase or D-mannitol. Catalase diminished the genotoxic activity of the mixture of ascorbic acid with copper ions in E. coli strains PQ37 and PQ 300, but genotoxicity of this mixture was not suppressed by superoxide dismutase and D-mannitol. The results suggest that copper ions diminished the content of peroxide generated by ascorbic acid.