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F.J. Wiebel T. Wolff M. Lambiotte 《Biochemical and biophysical research communications》1980,94(2):466-472
Cell lines derived from Reuber H-4-II-E hepatoma cells and their hybrids that differ in the expression of liver-specific functions are shown to contain different forms of monooxygenases. According to 1) the specificity toward the substrates benzo(a)pyrene, aldrin and chenodexycholic acid, 2) the kinetics of the epoxidation of aldrin, 3) the response to inducers, such as benz(a)anthracene and dexamethasone, and 4) the modifier 7,8-benzoflavone, the monooxygenases predominating in differentiated cell lines belong to the cytochrome P-450-dependent enzyme(s), those in the less differentiated lines belong to the cytochrome P-448-dependent form(s). 相似文献
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Quini CC Américo MF Corá LA Calabresi MF Alvarez M Oliveira RB Miranda JR 《Journal of biological engineering》2012,6(1):6-6
ABSTRACT: AC Biosusceptometry (ACB) was previously employed towards recording gastrointestinal motility. Our data show a reliable and successful evaluation of gastrointestinal transit of liquid and solid meals in rats, considering the methods scarcity and number of experiments needed to endorsement of drugs and medicinal plants. ACB permits real time and simultaneous experiments using the same animal, preserving the physiological conditions employing both meals with simplicity and accuracy. 相似文献
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We have examined the suitability of the continuous rat hepatoma cell line 2sFou for testing the genotoxicity of chemicals in comparison with that of primary rat hepatocyte cultures (HPC). The capacity of the cells for metabolic activation was assessed by measuring induction of DNA-repair synthesis and inhibition of replicative DNA synthesis by the test compounds dimethylnitrosamine (DMN), diethylnitrosamine (DEN), hydroxyurea (HU) and benzo[a]pyrene (BaP), which are substrates for major hepatic and extrahepatic forms of cytochrome P-450 dependent monooxygenases. The cellular capacity for DNA-repair synthesis was assessed using UV-light as a DNA-damaging agent. Repair-specific incorporation of [3H]deoxycytidine (3H-dCyd) caused by UV-light was higher in 2sFou cells than in HPC. In contrast, background repair incorporation of 3H-dCyd in 2sFou cells was only 1/3 that found in HPC. All the test agents induced DNA repair and inhibited DNA synthesis in both 2sFou cells and HPC. The two nitrosamines were more effective in HPC than in 2sFou cells. HU and BaP affected DNA repair and DNA synthesis in the two cell systems at a similar range of concentrations. In general, DNA repair in the 2sFou cells increased near linearly with the concentrations of the test compounds. The data indicate that 2sFou cells are capable of activating hepatotropic pro-mutagens/carcinogens such as dialkylnitrosamines, and are sensitive indicators of DNA damage. In contrast, BaP, a non-hepatotoxic compound, caused only little DNA repair in these cells. Thus, continuously growing cells, such as 2sFou, show a qualitatively similar response to genotoxic chemicals as HPC and offer a potential alternative to HPC for genotoxicity testing. 相似文献
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- 1 Ehrlich ascites tumor cells collected from donor mice on the 5th day after inoculation were injected into the peritoneal cavity of new recipient mice.
- 2 Cell cycle times were drastically shortened by transplantation, for instance, the length of the cell cycle from 47 to 21.5 hr, and the duration of S from 26.5 to 16.5 hr.
- 3 Transplantation also caused a transient delay of cells in G2 followed by a rapid acceleration and produced an immediate increase in the number of cells in DNA synthesis by about 5–8%.
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PAS1, a yeast gene required for peroxisome biogenesis, encodes a member of a novel family of putative ATPases 总被引:53,自引:0,他引:53
PAS genes are required for peroxisome biogenesis in the yeast S. cerevisiae. Here we describe the cloning, sequencing, and characterization of the PAS1 gene. Its gene product, Pas1p, has been identified as a rather hydrophilic 117 kd polypeptide. The predicted Pas1p sequence contains two putative ATP-binding sites and reveals a structural relationship to three other groups of proteins associated with different biological processes such as vesicle-mediated protein transport (NSF and Sec18p), control of cell cycle (Cdc48p, VCP, and p97-ATPase), and modulation of gene expression of the human immunodeficiency virus (TBP-1). The proteins share a highly conserved domain of about 185 amino acids including a consensus sequence for ATP binding. We suggest that these proteins are members of a novel family of putative ATPases and may be descendants of one common ancestor. 相似文献
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Laurent Corcos Jean-Pierre Rousset Franz Kiefer Friedrich J. Wiebel Mary C. Weisst 《Molecular & general genetics : MGG》1990,222(2-3):291-296
Summary We present a strategy to elucidate the rate-limiting steps in activation of carcinogenic compounds by cytochromes P450. The principle was to select Reuber rat hepatoma cells for resistance to a procarcinogen. The hypothesis was that resistant cells should be systematically deficient in the P450 enzyme(s) involved in the activation process. Here we present an example of the use of this approach using aflatoxin B1 (AFB1), a potent hepatocarcinogen, as the selective agent. Parental cells as well as individual and pooled colonies selected for AFB1 resistance from three independent rat hepatoma lines were characterized for their content of 1) mRNA hybridizing to cDNA and/or oligonucleotide probes for cytochromes P450IIB1, P450IIB2 and albumin; and 2) aldrin epoxidase activity. Parental aflatoxin B1-sensitive cells were shown to express P450IIB1 but not P450IIB2. The majority of the aflatoxin B1-resistant clones failed to accumulate cytochrome P450IIB1 mRNA and expressed no or only very low aldrin epoxidase activity. Albumin mRNA levels remained unchanged, demonstrating that loss of expression of cytochrome P450IIB1 was not a consequence of a general dedifferentiation event. A revertant population showing restoration of both cytochrome P450IIB1 mRNA accumulation and aldrin epoxidase activity was fully sensitive to aflatoxin B1. The correlation between expression of cytochrome P450IIB1 and sensitivity to aflatoxin B1 in both parental cells and revertants strongly suggests that cytochrome P450IIB1 is a major contributor to the activation of aflatoxin B1 in rat hepatoma cells. The kind of strategy described here could be applied to other compounds that become cytotoxic for hepatoma cells following activation by cytochromes P450.Abbreviations AFB1
aflatoxin B1
- AE
aldrin epoxidase
- AHH
aryl hydrocarbon hydroxylase
- PAH
polycyclic aromatic hydrocarbons
- PB
phenobarbital 相似文献
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Summary Polycyclic aromatic hydrocarbon-inducible monooxygenase directed toward the substrate benzo(a)pyrene, i.e., aryl hydrocarbon hydroxylase, was monitored in cell hybrids formed from mouse RAG cells and several human fibroblasts lines. In RAG cells no aryl hydrocarbon hydroxylase activity was detectable; however, these cells exhibited relatively high levels of NADPH cytochrome C (P-450) reductase (EC. 1.6.2.4). In 12 hybrids lines, induced aryl hydrocarbon hydroxylase segregated with human chromosome 2. The results indicate that the structural gene of the polycyclic aromatic hydrocarbon-inducible monooxygenase or gene(s) involved in the induction of the enzyme is located on human chromosome 2.Abbreviations AHH
aryl hydrocarbon hydroxylase
- IDH
isocitrate dehydrogenase
- MDH
malate dehydrogenase
- PAH
polycyclic aromatic hydrocarbons 相似文献
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