Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0019204 (hepatocellular carcinoma)
 71,386 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Expression of the ribose-positive phenotype was examined in hybrids obtained from the fusion of parental pentose-negative Novikoff hepatoma cells and ribose-positive variants. The two ribose-positive variants used differed phenotypically in their ability to use pentoses other than ribose for growth. One variant used D-ribose, D-xylose, and L-arabinose for growth, while the other variant used only D-ribose. Each variant was fused to pentose-negative parental hepatoma cells, and resultant hybrids were tested for the ability to use ribose. In both instances extinction of ribose utilization was the primary event, suggesting the existence of a trans-acting negative control element in the parental cells. In addition, hybrids from both fusion experiments eventually reexpressed the ribose phenotype. The rate of reexpression, however, was different for the two fusion experiments. Reexpression of ribose utilization in hybrids derived from the nonspecific variant occurred at approximately 10(-3) segregants/cell/day. Reexpressing segregants arose from the specific-derived hybrids at a rate of 0.5 segregants/cell/day. Possible reasons for this difference include a differential rate in chromosomal segregation or a difference in the regulation of ribose metabolism.
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PMID:Extinction and expression of the ribose-positive phenotype in hybrid Novikoff hepatoma cells. 679 62

Mouse teratocarcinoma cells (OTT6050) deficient for thymidine kinase were fused with rat hepatoma cells ( Fu5AH ) deficient for hypoxanthine phosphoribosyltransferase using inactivated Sendai virus. The hybrid cells were selected and cultured in the presence of HAT medium. A clonally established hybrid cell line ( As3 ), which in addition to its mouse genome contains several rat chromosomes, expresses rat specific enzyme variants and produces large primarily undifferentiated tumors, with some hepatoma characteristics in athymic nude mice. To reveal the in vivo developmental potential of these cells and to determine whether, under different experimental conditions, they are capable of participating in tissue differentiation, the As3 cells were injected into mouse blastocysts from the C57BL/6 strain. The experimental blastocysts were then transferred into the uteri of pseudopregnant foster mothers to allow further development. From a total of 212 blastocysts transplanted, 61 fetuses developed and were analysed for As3 contributions between the 10th and 18th day of gestation. Four fetuses at day 18 showed hybrid cell participation in their livers and a few organs of only endo-mesodermal origin, as judged from the presence of rat-specific enzyme variants. The enzymes were organ-specifically expressed (e.g., lactate dehydrogenase) or appeared newly during in situ differentiation while being absent in the original hybrid cells (e.g., glycerol-3-phosphate dehydrogenase). During short in vitro culture of the chimaeric organs, it was possible to select for the hybrid cells which reverted to an enzyme pattern simiar to but not identical with the As3 cell line and different to that observed in situ.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Tissue preference and differentiation of malignant rat x mouse hybrid cells in chimaeric mouse fetuses. 718 53

Pigmented subtetraploid subhexaploid mouse melanoma cells were fused with a range of different cell types. Expression of pigment formation appeared to be dependent on the phenotype of the non-melanoma parent cell, so that hybrids with lymphoid cells or chick embryo erythrocytes produced pigment, but hybrids between fibroblasts or epithelial rat hepatoma cells did not. The results were independent of gene dosage of either parent cell. gamma-irradiation of suppressing partner cells prior to fusion caused progressive increase in pigmentation with increasing dose of radiation. Cybrids between cytoplasts of suppressing fibroblasts and melanoma cells were pigmented.
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PMID:Persistence, suppression and re-expression of pigment formation in somatic cell hybrids between mouse melanoma cells and non-melanoma cells. 726 79

Enucleated chloramphenicol (CAP) resistant mouse L-cells (LEA-2A) were fused with the mouse hepatoma cells (BW1J). The resultant cybrids expressed CAP resistance (the property used in selection of the cybrids), and also expressed the hepatic-specific functions of the BW1J parent. Hybrids between these same cells, on the other hand, exhibited chloramphenicol resistance and extinction of the hepatoma-specific properties. Cybrids were also prepared between enucleated rat hepatoma cells (FT-2) and mouse erythroleukemia cells (C19TK). The resultant cybrids selected in tyrosine-free medium expressed phenylalanine hydroxylase, an enzyme normally appeared to be the result of activation of the previously silent gene of the C19TK cells. These cybrids, however, did not express any other liver-specific functions present in the FT-2 cytoplast donor. These experiments suggest that the transfer of heritable properties by cell cybridization is selection specific and that activation or extinction observed in hybrids may not occur in cybrids of the same cells.
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PMID:Transfer of heritable properties by cell hybridization: specificity and the role of selective pressure. 729 57

Phospholipids, glucolipids, and total proteins were separated from a plasma membrane fraction of rat liver. Membrane glycoproteins were isolated from deoxycholate extracts of rat liver membranes and hepatoma tissue culture membranes by concanavalin A chromatography. The membrane glycoprotein on hepatocytes that acts as a receptor for serum glycoproteins have lost their terminal sialic acid was also purified from rat liver membranes. Closed membrane vesicles were reconstituted from mixtures of deoxycholate-solubilized phospholipids and proteins by dialysis and purified by isopycnic centrifugation. The orientation of the proteins and glycoproteins in these reconstituted vesicles was examined by their accessibility to trypsin and neuraminidase and by their ability to be released from the vesicle by different concentrations of detergent. Most of the proteins are embedded in a right-side-out orientation in the lipid bilayer. The reconstituted membrane vesicles can be fused to mouse L-cells with polyethylene glycol. The extent of fusion is a function of the phospholipid:protein ratio in the reconstituted vesicles. After fusion, the phospholipid component of the vesicles mixes relatively rapidly with cell membrane lipids as judged by the immunofluorescence pattern of cells fused with lipid vesicles containing trinitrophenylated lipids. In contrast, proteins transferred to L-cells show restricted diffusion as judged again by immunofluorescence techniques. The metabolic turnover of proteins and glycoproteins after transfer to the plasma membranes of mouse L-cells was examined by radioisotopic methods. Total rat liver membrane proteins are very stable after transfer to the L-cells. Some of these proteins may be involved in the formation of an exoskeleton at the cell surface. Hepatoma tissue culture cell glycoproteins after transfer to the L-cells are less stable in terms of turnover properties than are total liver membrane proteins. However, some of these proteins are released into the medium as large molecular weight material rather than being degraded to small molecular weight, acid-soluble component. The receptor for serum asialoglycoproteins is relatively stable after transfer in reconstituted vesicles to the membrane of L-cells. Most of this hepatocyte-specific membrane glycoprotein is degraded to acid-soluble material with a half-life in the L-cell of at least 50 h. Transfer of the purified receptor in reconstituted vesicles to L-cells confers upon the recipient cell the biological activities specified and initiated by these receptors in hepatocytes.
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PMID:Insertion of biologically active membrane proteins from rat liver into the plasma membrane of mouse fibroblasts. 743 81

Fifty-five clones encoding epitopes of HCV were isolated from Japanese patients. Their amino acid homology (AAH) to the sequence of prototype (HCV-1) ranged from 47% to 94%. These sequences cover 60% of the HCV genome lacking M/E and NS2 regions suggesting a very low or lacking immunogenicity for these regions. Two test kits for detection of anti-HCV antibody were developed using a combination of a synthetic peptide (AR142) containing the epitope of N14 (QRKTKRSTNRR) having a homology to the core of HCV of 8/11AA and a non-fusion recombinant protein Y19 starting from amino acid number (AAN) 1380 to 1507 in the NS3 region showing a AAH to the HCV-1 of 90%, and a combination of a mixture of three synthetic peptides of S29 AAN of 1-30, 38-65 and 47-74 of the core and a non-fused recombinant protein S4 AAN of 1287-1506 having a 93% AAH of the NS3 region. They showed almost the same order of sensitivity and specificity of the second-generation kits when tested with serum from blood donors and patients with non-A, non-B hepatitis. It should also be stressed that in all of the complete responders of a recombinant alpha-interferon therapy, the antibody levels against AR142 gradually decreased during and after the treatment. In 1992, studies performed for 125 patients with hepatocellular carcinoma in our clinic shows that of these 16 patients might developed from either chronic non-B, non-C liver diseases or chronic liver diseases caused by mutant(s) of HCV as their serum were negative for HBsAg and second-generation of anti-HCV.
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PMID:Molecular cloning of HCV and clinical application. 752 19

Mouse F9 teratocarcinoma cells provide a system to study developmentally regulated alpha-fetoprotein (AFP) gene expression. AFP is not expressed in undifferentiated F9 cells but is induced when cells differentiate as cell aggregates in the presence of retinoic acid. Previous studies have led to the suggestion that undifferentiated F9 cells contain negative regulators of AFP expression. To test this, we have used transient heterokaryons to ask whether inactive AFP genes in undifferentiated F9 cells are responsive to positively acting trans-acting factors. Our results indicate that silent endogenous and transfected AFP genes are activated when undifferentiated F9 cells are fused to human hepatoma HepG2 cells. This suggests that the lack of AFP expression in undifferentiated F9 cells is due to the absence or insufficient level of positive-acting transcription factors, rather than the presence of dominant negative regulators. We also demonstrate that stably transfected AFP genes, although unmethylated, are properly regulated in F9 cells.
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PMID:Endogenous and transfected mouse alpha-fetoprotein genes in undifferentiated F9 cells are activated in transient heterokaryons. 754 61

A plasmid expression system has been developed which allows sequence-specific effects on mRNA degradation rates to be determined. This system uses stable, nonintegrating vectors that provide consistent levels of mRNA expression without the position effects common to integrating vectors. cDNAs encoding putative instability elements may be subcloned into the 5' untranslated region (5'UTR), the coding region, or the proximal 3'UTR of a beta-globin cDNA reporter. The effects of these sequences on mRNA stability may then be determined by actinomycin time course analyses of the fusion mRNAs and recombinant beta-globin mRNA in human cell lines. To demonstrate the utility of the vector system we fused an 820-bp fragment of the cDNA encoding the proximal 3'UTR of human 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase to the 3'UTR of the beta-globin reporter and introduced the vector into the human hepatocarcinoma cell line, HepG2. The fusion mRNA was degraded at a rate 2- to 2.5-fold greater than that of beta-globin alone, at a rate similar to that reported for HMG CoA reductase mRNA in normal rat liver. Similar to a number of other relatively unstable mRNAs, the rate of fusion mRNA degradation was greatly decreased by treatment with cycloheximide.
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PMID:An episomal expression vector system for monitoring sequence-specific effects on mRNA stability in human cell lines. 756 67

Cultured adult rodent hepatocytes are extensively used as a model system for gene transfer in vitro. In the present study, we examined the influence differentiation status and growth capacity of the hepatocytes on their infectivity in vitro by a retroviral vector. These parameters were initially studied in primary cultures of rat hepatocytes transduced with an ecotropic retroviral vector containing Escherichia coli beta-galactosidase. However, significant differences observed in the infectivity of hepatocytes from 12-day-old and adult rats led us to also examine hepatocytes from a transgenic mouse strain in which the SV40 large T antigen is fused to the regulatory sequences of the human anti-thrombin III gene. The large T antigen is expressed in the liver and these mice develop hepatoma within 7 months. A comparison of infectivity of hepatocytes from normal and transgenic mice of different ages indicated that in contrast to previous reports, hepatocytes which express differentiated functions during the first week of culture can still be efficiently infected by retroviral vectors. Optimal infection was observed between the second and fourth day of culture and does not appear to be due to transient cell dedifferentiation, but is more likely due to transient mitotic activity of mice cells since the role of growth factors seems crucial for infection. The peak of infection did not appear to correspond to transient cell dedifferentiation. We also found differences of infectivity between hepatocytes from normal and transgenic mice of different ages. Such differences are correlated with differences in in vitro BrdU incorporation, which was used to determine the proportion of dividing hepatocytes. These results indicate that the efficiency of infectivity of hepatocytes by recombinant retrovirus is probably related to their normal proliferative potential and not to some dedifferentiated stage. Hence these findings provide a model for efficient gene transfer in differentiated cells and suggest an approach for studies of liver-specific gene regulation and for somatic gene therapy of metabolic diseases as well.
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PMID:Retroviral infection of primary hepatocytes from normal mice and mice transgenic for SV40 large T antigen. 768 Oct 9

The P450/6 beta A (CYP3A2) gene encoding a testosterone 6 beta-hydroxylase is expressed predominantly in liver and induced by the treatment of rats with various compounds. To understand the mechanism of the basal transcriptional activation of the CYP3A2 gene, the cis-acting elements in the proximal promoter region (-165 to -73) of the CYP3A2 gene were identified in this study. Nuclear extract from rat livers interacted with three sites, 6 beta A-A (-106 to -87), 6 beta A-B (-140 to -119) and 6 beta A-C (-163 to -145). These sites were detectable by DNase I footprinting and gel mobility shift assays and found to share nucleotide sequence similarity with each other (T(A/C)(A/C)N(A/G)AAG(G/T)(C/T)CA). Direct repeats of AGTTCA (-134 to -120) and AG(G/C)TCA (-162 to -148) are also detected in 6 beta A-B and 6 beta A-C sites, respectively. To elucidate the relationship of these sites with basal transcriptional activation of the CYP3A2 gene, varying lengths of the proximal promoter region (-164 to +41) fused to a CAT reporter gene were transfected in human hepatoma (HepG2) and mouse adrenal tumor (Y-1) cells. The relative level of CAT activity in HepG2 cells was slightly increased by the deletion of the 5'-portion from -164 to -111 bp, but was reduced to 14% of the control (the construct including from -110 to +41) by the deletion from -110 to -81 including the 6 beta A-A site. On the other hand, these deletions have no clear effect on the level of the activity in Y-1 cells. Substitution mutations at two nucleotides in the 6 beta A-A site resulted in the reduction of CAT activity in HepG2 cells to 12% of the activity in the wild-type construct. The interaction of an oligonucleotide corresponding to the 6 beta A-A site (-106 to -87) with liver nuclear factors was completely inhibited by the addition of a typical oligonucleotide for hepatocyte nuclear factor-4 (HNF-4) binding site (F. M. Sladek, W. Zhong, E. Lai, and J. E. Darnell, Jr., 1990, Genes Dev. 4, 2353-2365) but not of oligonucleotides corresponding to 6 beta A-B or 6 beta A-C sites. These results suggest an essential role of the binding of HNF-4 and/or HNF-4-related nuclear factors to the 6 beta A-A site on the basal transcriptional activation of the CYP3A2 gene in liver cells.
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PMID:Transcriptional elements directing a liver-specific expression of P450/6 beta A (CYP3A2) gene-encoding testosterone 6 beta-hydroxylase. 772 76


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