Cell death induced in L-cells by treatment with thymidine: Staging of the process and relationship to apoptosis

Amenta, Joseph S.; Sargus, M. J.; Baccino, Francesco M.; Sacchi, Cristina; Bonelli, Gabriella

doi:10.1007/bf02631363

Cited by 7 publications

(3 citation statements)

References 22 publications

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“…Suppression of DNA and RNA synthesis (Parekh and Chitins, 1991) or inhibition of HMGCoA reductase (Castillo et al, 1990;Haughom and Spydevold, 1992) might well play a role, since apoptosis may be caused by inhibitors of macromolecular syntheses (e.g. Hickman, 1992;Amenta et al, 1993;Sherwood and Schimke, 1994;Barbiero et al, 1995) or by the HMGCoA reductase inhibitor lovastatin (e.g. Bansal et al, 1991;Pe Â rez-Sala and Molinedo, 1994;Han and Wyche, 1996).…”

Section: Mechanisms Of the Lethal Action Of Clofibrate And Nafenopinmentioning

confidence: 99%

Rapid and extensive lethal action of clofibrate on hepatoma cells in vitro

et al. 1997

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Clofibrate, for a long time in use as a hypolipidemic drug, is a well known peroxisomal proliferator (PP) and hepatocarcinogen in rodents. We show here that in vitro 1 mM clofibrate induces a rapid and massive death of rat AH-130 hepatoma cells. Cell death was prominent already after 4 h of treatment, with a characteristic`apoptotic' pattern by conventional microscopy. This was further supported by the pronounced chromatin condensation detectable on 4',6-diamine-2'-phenylindole dihydrochloride (DAPI) staining, the clearcut internucleosomal DNA fragmentation on agarose-gel electrophoresis (ladder patten), and the accumulation of markedly hypochromic cells observed in flow cytometric DNA histograms. Consistently with the apoptotic features of the process, some parameters commonly used to detect cell death, such as plasma membrane permeabilization to trypan blue or propidium iodide, lack of mitochondrial retention of rhodamine 123, or extracellular release of lactate dehydrogenase, were all virtually negative. However, these same parameters became markedly positive after 24 h of treatment, which was suggestive for the occurrence of`secondary' necrosis among AH-130 cells. By a combination of flow cytometric parameters, after 4 h on 1 mM clofibrate only 41% of the AH-130 cells could still be categorized as viable (i.e., non-apoptotic and non-necrotic), while 46% of cells appeared apoptotic and 13% necrotic. At 24 h, 67% of cells were necrotic, 20% apoptotic and only 13% non-apoptotic and nonnecrotic. Apoptosis was also extensive in AH-130 cells treated with another PP such as nafenopin at 1 mM concentration and in human hepatoma HepG2 cells treated with clofibrate. By contrast, clofibrate did not cause apoptosis on primary rat hepatocyte cultures. These observations indicate that: (i) apart from their well-known cell growth-promoting action, PPs such as clofibrate or nafenopin may exert a substantial cytotoxic action on targets such as the AH-130 or HepG2 hepatoma cells; (ii) this cell death evolves from an initial apoptotic' to an eventual`necrotic' pattern; (iii) detection of cell death requires the adoption of a full panel of tests, adequate to cover the whole evolving death pattern, while such tests may even be substantially misleading whenever applied individually; (iv) the cytotoxicity of clofibrate and similar agents on normal and, particularly, tumoural cells may deserve careful reevaluation.

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Section: Mechanisms Of the Lethal Action Of Clofibrate And Nafenopinmentioning

confidence: 99%

Rapid and extensive lethal action of clofibrate on hepatoma cells in vitro

et al. 1997

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“…In vitro assays with osteoblastic cells are a useful method for evaluating the interactions of biomaterials and osteoblastic cells. 3 H-thymidine incorporation into DNA, detected by autoradiography, has been widely applied in cell proliferation studies (Amenta et al 1988(Amenta et al , 1993Bilir et al 2002). The main drawbacks of this technique are potentially hazardous radioactivity and the time required for development of autoradiographs.…”

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confidence: 99%

Biocompatibility of different barrier membranes in cultures of human CRL 11372 osteoblast‐like cells: an immunohistochemical study

Bılır

Aybar

Tanrikulu

et al. 2006

Clinical Oral Implants Res

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“…In this study we utilized exponentially growing L-cells (ATCC American Type Culture Collection, Rockville, MD, USA) derived from mouse fibroblasts (Amenta et al 1988(Amenta et al , 1993Ridvanogullari et al 1993). Cells were maintained in Medium 199 M199 (Biological Industries (BI), Haemek, Israel) (M199) supplemented with 10% fetal bovine serum, 100 mg/ml penicillin, 100 mg/ml streptomycin in 25 cm 2 flasks.…”

Section: Cell Culturementioning

confidence: 99%

The effects of naproxen sodium on specific parameters of L‐strain cells over guided bone regeneration barrier materials

Aybar

Erkut

Yıldırım

et al. 2004

Clinical Oral Implants Res

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There is strong evidence that nonsteroidal antiinflammatory drugs (NSAID) may exert a significant antiproliferative effect. This study evaluated the influence of NSAID on specific parameters of fibroblastic cells, in vitro, over two-guided bone regeneration (GBR) barrier materials. Fibroblast cells were cultured on bioabsorbable membrane made of collagen (Bio-Gide(R)- BG) and the most common nonresorbable membrane which is made of expanded polytetrafluoroethylene (ePTFE, Gore-Tex(R)- GT). Naproxen sodium (10 mM) was used as an analgesic drug. The fibroblast cells were cultured in vitro for 24 h and examined by scanning electron microscopy (SEM). Cells were cultured in the presence of (3)H-thymidine to study cell proliferation. And also cell numbers and viabilities were measured. The difference between the means for each group were analyzed for statistical significance by Kruskal-Wallis one-way ANOVA followed by post hoc comparisons using the Dunn statistical method. Of all the six groups, the control group stimulated DNA synthesis more than the others. With respect to cell numbers, there was statistically significant difference between the control group and naproxen planted BG membrane group. The interpretation of our SEM images is that these two barriers and naproxen seem to have had the least effect on cellular morphology. These data suggest naproxen have an inhibitory effect on stimulation of DNA synthesis, cell numbers and viabilities. And also lacking adherence of cells to the membranes may be due to the physical properties of the materials.

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Cell death induced in L-cells by treatment with thymidine: Staging of the process and relationship to apoptosis

Cited by 7 publications

References 22 publications

Rapid and extensive lethal action of clofibrate on hepatoma cells in vitro

Rapid and extensive lethal action of clofibrate on hepatoma cells in vitro

Biocompatibility of different barrier membranes in cultures of human CRL 11372 osteoblast‐like cells: an immunohistochemical study

The effects of naproxen sodium on specific parameters of L‐strain cells over guided bone regeneration barrier materials

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