The metabolism of beef tallow sucrose esters in rat and man.
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Adipose Tissue↗
Biomedical subjects
C J Marshall
Publications and source records attributed to C J Marshall.
Expression of the transformed phenotype and tumorigenicity in somatic cell hybrids.
We have previously shown by examining the anchorage dependence, density-dependent inhibition of growth, LETS protein and microfilament bundles that the transformed phenotype of the parental tumours are suppressed in hybrids between rat embryo fibroblast (REF) and mouse tumour cells (TA3B). Hybrids between TA3B and Syrian hamster sarcoma cells (BHK-B1) also show suppression. We now demonstrate that tumorigenicity in nude mice is also suppressed in TA3B X REF and B1 X TA3B hybrids. Tumours arise from the suppressed hybrids by the selective outgrowth of variants with properties different from the majority of cells inoculated. These tumour variants always had an altered cytoskeletin but 1 out of 14 cases retained anchorage-dependent growth. Selection in culture for anchorage-independent growth selected for tumorigenicity. The coordinate suppression of the transformed phenotype and tumorigenicity could be explained by postulating a pleiotropic control mechanism affecting both. However, the occasional dissociation of tumorigenicity from anchorage dependence in the variants suggests that the targets for the control mechanism must be different.
Animals↗
Ruthenium red-positive surface layer, extracellular filamentous material and intercellular junctions in hybrids between tumour and normal cells: abundant gap junctions correlate with density-dependent inhibition of growth.
The cell surface and intercellular junctions of transformed and non-transformed hybrid cells, obtained from fusion of murine mammary adenocarcinoma cells (TA3B) and normal rat embryofibroblasts (REF), were compared with those of the parental cells, using ruthenium red (RR) staining of cells fixed in situ. An RR-positive layer of variable thickness was found on the surface of all cell types. Measurements of the thickness of this layer on the free surface of the cell cultures deomonstrated a significant difference between transformed and non-transformed cells. The thickness distribution of the RR layer was similar on the surface of REF and non-transformed hybrid cells, but there was significant variation among all the transformed cell lines. Extracellular filamentous RR-positive material, usually in direct contact with the cell surface, was present in confluent cultures of REF and non-transformed hybrid cells, but was absent in the transformed cell lines. Gap junctions were few in both parental cell lines and rare or absent in transformed hybrids; a large increase in number and size of gap junctions was found in non-transformed hybrids. Abundance of long gap junctions was correlated with density-dependent inhibition of growth, since non-transformed hybrids grew in single layers whereas both normal and tumour parental cell types, and transformed hybrids grew in multilayers. Tight junctions were frequently encountered in TA3B and transformed hybrid cells but were not seen in REF cells and only occasionally seen in non-transformed hybrids. Intermediate-type junctions occurred in all cells, but desmosomes were found only rarely in TA3B cells and never in the other cell types.
Adenocarcinoma↗
Suppression of the transformed phenotype in somatic cell hybrids.
Somatic cell hybrids between mouse mammary tumour cells (TA3B) and diploid rat embryo fibroblasts (REF) or between TA3B and Syrian hamster sarcoma cells (BI) were examined for the in vitro characteristics of transformed cells as soon as possible after cell fusion. Unlike the parental tumour cells as three of four TA3B X REF and five BI X TA3B independent hybrid lines had low colony-forming efficiencies in agar, exhibited density-dependent inhibition of growth and did not form colonies on confluent monolayers of 3T3 cells, demonstrating that the transformed phenotype was suppressed in these hybrids. In addition tests of some of the hybrid lines for tumour production in nude mice showed that this was also suppressed. Suppression was more stable in the TA3B X REF than in the BI X TA3B hybrids, variants of the BI X TA3B hybrids with the properties of transformed cells could be readily isolated by subculturing cells that had grown in agar. Tumour growth selected for hybrids with the characteristics of transformed cells, and derivatives of the hybrids selected to show the transformed phenotype readily produced tumours. These correlations suggest that the transformed phenotype and malignancy may be under the same control in these cells. The phenomenon of suppression may be explained by the hypothesis that neoplastic transformation results from recessive mutations in genes which control the normal phenotype. On this model the finding of suppression in hybrids between two different tumour lines is interpreted as complementation and indicates that the mutations are not the same in all cell lines.
Animals↗
Microfilament bundles, LETS protein and growth control in somatic cell hybrids.
Hybrid cell lines between normal rat embryo fibroblasts and TA3B mouse tumour cells, or between TA3B and BI hamster sarcoma cells, have been examined for the expression of the cell surface large external-transformation-sensitive (LETS) protein and the organization of microfilament bundles. LETS protein was detected by lactoperoxidase-catalysed radioiodination and microfilament bundles were visualized by indirect immunofluorescence with antibodies directed against actin or myosin. Hybrids which exhibited normal growth control characteristics had high levels of LETS protein and extensive microfilament bundles. Neoplastic transformation appears to be suppressed in these hybrids. Hybrids which had the growth control characteristics typical of transformed cells had reduced or zero levels of LETS protein and few microfilament bundles. These results confirm previous studies on the expression of the transformed phenotype in these hybrids and demonstrate that there is a good correlation between normal growth control and the presence of LETS protein and microfilament bundles. However, the changes in cell surface LETS protein and in the organization of microfilament bundles often appeared to be quantitative reductions rather than all-or-none effects. The magnitude of the alterations in the levels of LETS protein and in the organization of microfilaments appeared to correlate with the range of transformed characteristics exhibited by the cells. One transformed hybrid in particular, selected for growth in agar, had some surface LETS protein, some microfilament bundles and retained density-dependent inhibition of growth.
Cell Transformation, Neoplastic↗
Markers of neoplastic transformation in epithelial cell lines derived from human carcinomas.
Density-dependent inhibition of growth, plating efficiency on confluent monolayers of 3T3 cells, and growth in agar have been measured in epithelial tumor cell lines to determine whether they have properties in common with transformed mesenchymal cells. Five lines (RT4, RT112, J82, T24, and EJ) were derived from different human bladder tumors and HT29 was from a human colon tumor. All the lines resembled transformed "fibroblasts" in the absence of density-dependent inhibition of growth and the cell-surface large external transformation-sensitive protein, and they could form colonies on 3T3 monolayers. Only RT112, EJ, and HT29 were tumorigenic in nude mice, and the tumors had many of the structural and ultrastructural features of both the original tumor and the tissue of origin, even though the cells had been through many in vitro passages. Four of the lines (J82, T24, EJ, HT29) grew in agar, so that in some cell lines no correlation of growth in agar with tumorigenicity in nude mice was found: J82 and T24 were nontumorigenic and grew in agar, whereas tumorigenic line RT112 did not grow in agar. The ability to grow in agar did not appear associated with the production of high levels of plasminogen activators.
Animals↗
Chromosomally depleted interspecific hybrid cell clones selected with cytotoxic antisera: utilization in the study of control of murine leukemia virus host-range.
A chromosomally stable mouse-Chinese hamster hybrid cell line was subjected to five rounds of selection with cytotoxic antisera raised in rabbits against either the parental mouse 3T3 cells or the parental Chinese hamster Wg-1 cells. Routine karyological analysis of clones isolated at each stage of serum selection revealed that treatment with either serum resulted in a limited loss of chromosomes (compared to the untreated hybrid cell cultured in parallel) and that the pattern of chromosome loss could not be correlated with the particular antiserum used for selection. However, more detailed analysis with the SSC-formamide C-banding technique, which identifies chromosomes containing a mouse centromere region, demonstrated that while large-scale chromosome loss was not achieved as a result of antiserum selection, the limited loss of chromosomes did, in fact, reflect a specific depletion of chromosomes in response to treatment with cytotoxic antiserum. Specific chromosomal elimination was shown to occur as early as the first round of antiserum treatment. Antigenic analysis of the serum-selected clones revealed a quantitative decrease in the expression of the species-specific surface antigens selected against, but no qualitative loss of antigens was detected. The results suggest that treatment with cytotoxic antiserum may select for clones that have lost specific chromosomes bearing genes regulating the expression of species-specific surface antigens, rather than for those demonstrating large-scale depletion of chromosomes bearing the corresponding structural genes. Some of these chromosomally depleted hybrid cell clones have been used (along with pseudotype viruses containing the genome of vesicular stomatitis virus within the envelope of murine leukemia virus, VSV [MuLV]), to study the mechanisms regulating MuLV replication in Chinese hamster cells. The results indicate that the restriction of MuLV replication in Chinese hamster cells operates at two levels: (a) an inability to adsorb to or penetrate Chinese hamster cells; and (b) an additional intracellular block which is dominant in the mouse-Chinese hamster hybrid cell clones examined. This latter block is presently under study.
Antigens↗
A method for analysis of chromosomes in hybrid cells employing sequential G-banding and mouse specific D-banding.
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Animals↗
Selective depletion of chromosomes in a stable mouse-Chinese hamster hybrid cell line using antisera directed against species-specific cell surface antigens.
A chromosomally-stable mouse-Chinese hamster hybrid cell line was subjected to five rounds of selection with cytotoxic antisera raised in rabbits against either the parental mouse 3T3 cells or the parental Chinese hamster Wg-1 cells. Routine karyological analysis of clones isolated at each stage of serum selection revealed that treatment with either serum resulted in a limited loss of chromosomes (compared to the untreated hybrid cell cultured in parallel) and that the pattern of chromosome loss could not be correlated with the particular antiserum used for selection. However, more detailed analysis with the SSC-formamide C-banding technique, which identifies chromosomes containing a mouse centromere region, demonstrated that while large-scale chromosome loss was not achieved as a result of antiserum selection, the limited loss of chromosomes did, in fact, reflect a specific depletion of chromosomes in response to treatment with cytotoxic antiserum. Specific chromosomal elimination was shown to occur as early asthe first round of antiserum treatment. Antigenic analysis of the serum-selected clones revealed a quantitative decrease in the expression of the species-specific surface antigens selected against, but no qualitative loss of antigens was detected. The results suggest that treatment with cytotoxic antiserum may select for clones that have lost specific chromosomes bearing genes regulating the expression of species-specific surface antigens, rather than for those demonstrating large-scale depletion of chromosomes bearing the corresponding structural genes.
Antigens↗
Synthesis of ribosomal RNA in synkaryons and heterokaryons formed between human and rodent cells.
A study has been made of the ribosomal RNA and chromosome constitution of man-mouse hybrid cells. Previous work has shown that no human 28s rRNA is detectable in man-mouse synkaryons. In general human chromosomes are lost from such hybrids. With a recently developed method for distinguishing mouse from human chromosomes, an analysis of various man-mouse hybrid cell lines has been made. This indicates that not all the human chromosomes bearing nucleolar organizers are lost in the hybrid cells and such loss cannot alone explain the absence of human 28s rRNA. An examination of the 28s rRNA synthesized by heterokaryons formed from several different parent cells has revealed that both parental types of 28s rRNA are present in heterokaryons. The control of rRNA synthesis in hybrid cells is discussed.
Animals↗