Detection of an early surface change during oncogenic transformation.

Parry, Gordon; Hawkes, Susan P.

doi:10.1073/pnas.75.8.3703

Cited by 17 publications

(9 citation statements)

References 16 publications

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“…This possibility cannot be ruled out at this time. Previous studies with this particular clone of LA24 in our laboratory, however, have shown tight control at 410C for morphology, 2-dGlc uptake, and collagen synthesis in PAT cells (22), and fluorescamine labeling, dThd incorporation, morphology, and 2-dGlc uptake in CEF (29).…”

Section: Resultsmentioning

confidence: 99%

Is the product of the src gene a promoter?

Bissell¹,

Hatié²,

Calvin³

1979

Proc. Natl. Acad. Sci. U.S.A.

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Addition of a potent promotor, 12-O-tetradecanoylphorbol 13-acetate (TPA), to primary avian tendon or chicken embryo fibroblast cells infected with a temperaturesensitive mutant of Rous sarcoma virus produced a complete transformed phenocopy at the non rmissive temperature by the criteria tested. While normal, uninfected cultures also shifted towards a transformed phenotype after TPA addition, they did not achieve the same degree of morphological and biochemical alteration seen in virus-infected, TPAtreated cells.It is proposed that viral carcinogenesis, despite its rapidity, may occur in two stages: an "initiation" step caused by expression of a part of viral genome other than src (or by integration) and a promotion step (itself a multistep process) caused by the activation of the src gene. The src gene product could be enhanced or replaced by other promoting agents.We have gained tremendous knowledge of the structure and the genetics of RNA tumor viruses in recent years (1-3), yet we know relatively little about the mechanism(s) of viral carcinogenesis. The early optimism that viral transformation could provide a simple model for studying malignancy and that, once we identified the "oncogene" (4), we would understand cancer, has given way to skepticism and to an abrupt shift of interest from tumor viruses to other environmental carcinogens.The rapidity and completeness by which RNA tumor viruses could transform cells in culture andthe simplicity of the assay systems were the initial arguments for the use of RNA tumor virus-transformed cells as models for studying malignancy. These same reasonings, however, now are used to The isolation of active ingredients of croton oil by Hecker (7) has opened a whole new area of research, that of mechanism of action of tumor-promoting agents 8-11. It is now apparent that many characteristics induced by Rous sarcoma viruses (RSVs) are also induced by tumor promoters (10, 11). The overlap, however, is incomplete. Indeed, most of the effects of tumor promoters on normal cultured cells are analogousqualitatively and quantitatively-to the effects of serum (12) or other growth factors and are less pronounced than those caused by viral transformation. In addition, some characteristics, such as transformed morphology (11) and growth in agar, are not induced by the promoters.In this paper we show that at 410C addition of 12-0-tetradecanoylphorbol 13-acetate (TPA), a potent chemical promoter, to chicken cells infected with LA24, a temperature-sensitive (ts) mutant of RSV with a defect in the sarc gene, produces a more exact phenocopy of transformed cells than does TPA treatment of normal cells. The product of the arc gene at the nonpermissive temperature, therefore, appears to be either activated or replaced by the promoter. We propose that the product of the src gene itself may be a promoter rather than the initiator of transformation and that viral carcinogenesis may occur in two stages similar to those operating in mouse skin (13). The predictions of the model and the a...

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Section: Resultsmentioning

confidence: 99%

Is the product of the src gene a promoter?

Bissell¹,

Hatié²,

Calvin³

1979

Proc. Natl. Acad. Sci. U.S.A.

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“…Labeling of cells with fluorescamine and analysis of fluorescent products by SDS-polyacrylamide gel electrophoresis: This was carried out as previously described (18). Briefly, 2 ml of labeling solution consisting of 1 mg fluorescamine/0.5% acetone/0.2 Msodium borate, pH 9.0, was added to monolayers.…”

Section: Methodsmentioning

confidence: 99%

“…Alterations which occur as a result of oncogenic transformation have been characterized using a variety of techniques. One that we have developed and used extensively is the reaction of monolayers of cultured cells with fluorescamine (8,9,18,19), a compound that reacts with primary amines to form fluorescent adducts. We have demonstrated that primary and established lines of cells transformed by viruses and a chemical carcinogen exhibit three-to four-fold lower fluorescence intensities than do their nontransformed counterparts (7).…”

mentioning

confidence: 99%

“…The difference in reactivity towards fluorescamine is an early alteration during the development of the transformed phenotype as exhibited by chicken embryo fibroblasts infected with a temperature sensitive mutant of Rous sarcoma virus. It is more directly related to the transformation process than to the growth rate of the cells and is unrelated to the loss of surface proteins such as fibronectin (18).…”

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confidence: 99%

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Detection of transformed cells using a fluorescent probe: The molecular basis for the differential reaction of fluorescamine with normal and transformed cells

1982

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Normal and transformed fibroblasts can be discriminated by a flow cytometry assay on the basis of their differential reaction with fluorescamine. The cause of altered reactivity of transformed cells with this fluorescent probe has been investigated by a detailed analysis of its reaction with chicken embryo fibroblasts transformed by a temperature sensitive mutant of Rous sarcoma virus. The subcellular distribution of fluorescent adducts characterized by cell fractionation and gel electrophoresis procedures supports the hypothesis that transformed cells possess a surface barrier which decreases the accessibility of fluorescamine to reactive macromolecules. The barrier has been identified as being composed at least partly of hyaluronic acid, because of the ability of purified and specific hyaluronidase (from Streptomyces hyalurolyticus) to modulate the response of transformed cells to fluorescamine. Enzyme treatment of transformed cells prior to reaction with fluorescamine causes them to resemble nontransformed cells both in the nature of components labeled and in their fluorescence intensity. It is suggested that fluorescamine monitors an altered surface hyaluronic acid composition which occurs upon transformation. Its significance is discussed in terms of the known physical properties of the molecule and the finding that it is an early event in the process of transformation.

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“…There is the added advantage and flexibility of easily snapping the harness on and off during and after experimental periods, permitting recordings from a number of animals with the same transmitter. [33][34][35][36][37][38][39][40][41][42][43][44][45][46][47][48][49][50]1977), In subsequent experiments involving about 400 rats we have investigated how magnitude of remote loss varied with (a) cortical region, (b) size of lesion, (c) time elapsed between lesion and sacrifice, and (d) post-operative environment, either enriched (EC) or impoverished (IC). Somesthetic cortex, although adjacent to the occipital area, showed only about 2% loss of DNA while remaining dorsal cortex and ventral cortex lost 6-8%; hippocampus showed no change.…”

Detection of an early surface change during oncogenic transformation.

Cited by 17 publications

References 16 publications

Is the product of the src gene a promoter?

Is the product of the src gene a promoter?

Detection of transformed cells using a fluorescent probe: The molecular basis for the differential reaction of fluorescamine with normal and transformed cells

Chemical Biodynamics Division. Annual report 1979

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