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Biomedical subjects

H M Wallace

Publications and source records attributed to H M Wallace.

At least 19 recordsLinked to original sources
PubMed1994

Regulation of spermidine/spermine N1-acetyltransferase in human tumour cells.

It seems therefore that the induction of N1-SAT has a number of facets: some stabilization of the enzyme protein; stabilization of mRNA; requirement for new protein and RNA synthesis; and a Ca(2+)-sensitive induction site. It may be there is a polyamine-analogue-binding/induction site as well as an induction site which binds agents such as doxorubicin and butyrate (Figure 3). At present it is not clear whether the polyamine analogues interact directly with the active site of N1-SAT and it will be interesting to determine the effect of site-specific mutagenesis on the inducibility of N1-SAT by both polyamine analogues and antitumour agents like doxorubicin. Alternatively, it may be that agents such as doxorubicin and butyrate alter the conformation of DNA and thus release polyamines from intracellular binding sites. This increase in intracellular polyamine content may then induce N1-SAT activity which in turn will activate polyamine efflux with the net result being a decrease in intracellular polyamine content and the rate of cell growth.

Acetyltransferases↗
PubMed1992

Utilization of extracellular lipids by HT29/219 cancer cells in culture.

Uptake and incorporation of long-chain fatty acids were studied in a human colorectal cancer cell line (HT29/219) grown in culture medium supplemented with either fetal calf serum (FCS) or horse serum (HS). The cells were grown for 120 h with no change of medium; the two major cellular lipid classes, the phospholipids and the triacylglycerols, were analyzed at regular time-points. We observed significant changes in the concentration of most fatty acids throughout culture, and differences in their composition when different sera were used to supplement the medium. Minimal levels of free fatty acids were found in the cells, indicating a very small "free fatty acid pool". A major difference between the cells grown in media supplemented with different sera was the changes observed in concentrations of cellular polyunsaturated fatty acids during growth. In cells grown with FCS (in which 20:4n-6 is present), the levels of this acid in the phospholipid and triacylglycerol fractions declined rapidly during cell growth, suggesting further metabolism. In cells grown in medium supplemented with HS, 18:2n-6 was the major polyunsaturated acid present. There was clear evidence that this acid accumulated in the cellular triacylglycerol and phospholipid fractions. Furthermore, its concentration did not decline during growth in culture, suggesting minimal conversion to other polyunsaturated n-6 acids. Our results suggest that fatty acids from additional sources in the medium, for example triacylglycerols and phospholipids associated with the lipoproteins, are taken up by the cells. There is also indication of cellular fatty acid synthesis, particularly of monounsaturated and saturated acids during the culture period.(ABSTRACT TRUNCATED AT 250 WORDS)

Arachidonic Acid↗
PubMed1992

Effects of cyclosporin A and a non-immunosuppressive analogue, O-acetyl cyclosporin A, upon the growth of parent and multidrug resistant human lung cancer cells in vitro.

We have studied the ability of cyclosporin A (CsA) and a non-immunosuppressive analogue, O-acetyl cyclosporin A (OACsA, B3-243) to inhibit the growth of human lung cancer cells in vitro. Using continuous drug exposure and the MTT colorimetric assay to determine cell growth we found that CsA produced partial growth inhibition at doses ranging from 0.5 to 3.0 micrograms ml-1 (0.4-2.4 microM). At progressively higher doses, complete growth inhibition and in situ cell lysis were seen. The P-glycoprotein expressing multidrug resistant (MDR) variant H69/LX4 of the small cell line H69/P was less sensitive to cyclosporins than the parent line, but this was not true of the non-P-glycoprotein expressing MDR variants of large cell line COR-L23 or adenocarcinoma line MOR. Sensitivity to OACsA was approximately 2-fold higher than that to CsA in most of the lines although not in the most sensitive line, COR-L88. Even in COR-L88, exposed to CsA or OACsA for 24 h, clonogenic cell survival was reduced only to 50%. There was no reduction in polyamine content of COR-L23 or COR-L88 cells following 48 h of exposure to CsA or OACsA. The effects on cell growth could not be inhibited by the addition of exogenous putrescine, nor could they be enhanced by the addition of alpha-difluoromethylorthinine. It does not appear therefore that inhibition of polyamine synthesis is the basis of the observed growth inhibition.

Adenocarcinoma↗
PubMed1991

Mechanisms of spermine toxicity in baby-hamster kidney (BHK) cells. The role of amine oxidases and oxidative stress.

Spermine was toxic to BHK-21/C13 cells in the absence of any extracellular metabolism of the amine. Inhibition of copper-containing amine oxidases with aminoguanidine partially prevented the response, whereas inhibition of polyamine oxidase with MDL-72,527 exacerbated the effect. Oxidation by an intracellular copper-containing amine oxidase may be involved in the toxicity of spermine, whereas the polyamine-interconversion pathway appears to play a cytoprotective role. There was no evidence for spermine imposing a state of oxidative stress within the cells. Inhibition of catalase and glutathione reductase did not alter the cytotoxicity of spermine, and there was no excretion of oxidized glutathione into the extracellular medium. The results suggest that spermine itself can exert a toxic effect directly on the cells.

Animals↗
PubMed1991

Effects of cyclosporin A on growth and polyamine metabolism of MOLT-4 T-lymphoblastic leukaemia cells.

We have examined the effects of Cyclosporin A (CsA) on growth and polyamine metabolism of MOLT-4, human T lymphoblastic leukaemia cells to ascertain the role of the polyamine biosynthetic pathway in the antitumour action of CsA. We observed that CsA had a dose-dependent inhibitory effect on growth of the cells in vitro, decreasing protein content, cell number and the rate of incorporation of 3H-thymidine into the cells. However, CsA treatment had no significant effect on intracellular polyamine levels in the cells. Contrary to previous reports, simultaneous addition of the diamine, putrescine, with CsA did not block or lessen the growth inhibitory effects of CsA. On the other hand, ornithine decarboxylase activity, the rate limiting enzyme of polyamine biosynthesis which converts ornithine to putrescine, was decreased by CsA treatment. This decrease appeared to be reversible and contrasts with the inhibition by alpha-difluoromethyl-ornithine, which is irreversible and can be overcome by addition of putrescine. This suppression of ornithine decarboxylase by CsA is more likely to occur by indirect effects on translation and/or transcription rather than a direct effect on the enzyme. It may be a contributory factor in the overall antiproliferative effects of CsA but is more likely to be a response to these growth inhibitory effects rather than a direct effect of the drug.

Cell Division↗
PubMed1990

Spermine toxicity and glutathione depletion in BHK-21/C13 cells.

Spermine, a polycationic amine, produced a dose-dependent inhibition of BHK-21/C13 cell growth. This response was not due to the extracellular metabolism of spermine by an amine oxidase found in bovine serum, as the toxicity was observed when the cells were grown in medium supplemented with horse serum. Three indices were used to monitor cell growth, cell number, protein content and [3H]thymidine incorporation into DNA. Spermine (2mM) caused significant reductions in all three measurements after a 6-8 hr exposure. The amine was rapidly taken up into the cells reaching levels 15-16-fold greater than in control cells within 2 hr. There was a rapid loss of intracellular reduced glutathione following exposure to toxic concentrations of spermine, which occurred before any effect on cell growth. Three methods for the determination of intracellular glutathione content were compared in this system. The effect on both cell growth and glutathione was reversible following removal of spermine from the extracellular medium. The possible mechanisms involved in this toxic response are discussed with particular reference to the depletion in intracellular reduced glutathione.

Animals↗
PubMed1990

The potential of cyclosporin A as an anti-tumour agent.

Cyclosporin A (CsA) has become established as the agent of choice for the prevention of organ allograft rejection and has shown considerable promise in the clinical management of certain autoimmune disorders. The impact of CsA as an immunotherapeutic agent of major importance is attributable to its powerful, selective inhibitory action on T-lymphocyte activation and proliferation. Moreover, CsA lacks the myelotoxic and other major side effects associated with cytotoxic immunosuppressive agents, such as cyclophosphamide or azathioprine. It is now clear that CsA has a potential therapeutic role in the treatment of malignancies, especially T-cell cancers. Recent studies suggest that there may be several areas of application for CsA, either as a direct antiproliferative agent or in combination with other drugs, including inhibitors of polyamine biosynthesis or cytotoxic anti-tumour agents, including vincristine and adriamycin. In addition, CsA and non-immunosuppressive analogues have been shown to restore multi-drug sensitivity in cancer cells with acquired drug resistance. A further application of CsA may be to prevent the induction of human immune responses to therapeutic mouse monoclonal antibodies directed against tumour antigens, thereby enhancing the efficiency and safety of this form of cancer immunotherapy. Due to our incomplete understanding of the antiproliferative properties of CsA, further exploration of its potential as an anti-tumour agent must be accompanied by detailed studies aimed at elucidating its action on subcellular molecular events in both normal and malignant cells.

Animals↗
PubMed1990

Study of fatty acid profiles in cancer cells grown in culture using gas chromatography--mass spectrometry.

The determination of long-chain fatty acids in the phospholipid, triglyceride and free fatty acid fractions of HT29/219 colon cancer cells grown in a medium containing either foetal calf serum or horse serum, was carried out using gas chromatography - mass spectrometry. Several bonded-phase capillary columns were tested for the separation of the fatty acid methyl esters, and a 30-m poly(ethylene glycol) column was found to give optimum separation. The mass spectrometer was set to the multiple ion detection mode to increase the sensitivity of the recording of the characteristic ions, consisting of the molecular ion and the base peak. The phospholipid and triglyceride compositions of the cells were different when the cells were grown in media containing different sera. Differences were also found in the turnover of the acids in the different lipid fractions, the phospholipids being the most important, when the cells were grown in different media. The cellular metabolism and turnover of certain fatty acids differed from others in the same cell. These differences emphasise the importance of a precise knowledge of the lipid composition of the culture medium in in vitro studies of cancer cells.

Fatty Acids↗