Apoptosis is not involved in the hypersensitivity of fanconi anemia cells to mitomycin C

doi:10.1016/0165-4608(94)90218-6

Cancer Genetics and Cytogenetics

Volume 75, Issue 1, 1 July 1994, Pages 67-71

https://doi.org/10.1016/0165-4608(94)90218-6 Get rights and content

Abstract

A striking feature of Fanconi anemia (FA) cells is their hypersensitivity towards crosslinking agents such as mitomycin C (MMC). In this study, we have shown that treatment of lymphoblastoid cells with MMC resulted in nuclear fragmentation, chromatin condensation, and DNA degradation, which is characteristic of apoptosis. The level of DNA fragmentation 48 hours after MMC treatment reached approximately 33% in both control and FA cells. In addition, 24 hours after drug addition a decrease in the number of cells in the G2/M phase of the cell cycle was seen. This coincided with the appearance of apoptotic cells in the sub-G1 phase, indicating that once the cells had passed through G2/M, apoptosis occurred. The number of apoptotic cells increased to 60% 96 hours after MMC treatment. The number of apoptotic cells increased to 60% 96 hours after MMC treatment. The onset and level of apoptosis was found to be identical in FA and control cell lines, indicating that the FA defect does not lead to abnormal apoptotic cell death.

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Cited by (15)

The Fanconi anemia pathway and the DNA interstrand cross-links repair
2003, Biochimie
Fanconi anemia (FA) is a genetic cancer-predisposition syndrome characterized by bone marrow failure and cellular and chromosomal hypersensitivity to DNA cross-linking agents. Seven FA genes have been isolated and their products associate to form a pathway that interacts functionally or physically with several DNA-damage response proteins involved in cell cycle checkpoints and/or DNA repair. These proteins include BLM, ATM, BRCA1, XPF and the MRE11/RAD50/NBS1 complex. In spite of several recent striking progresses in the biochemistry and the molecular biology of the disorder, the precise function(s) of the FA proteins remain(s) poorly determined. However, several recent data indicate that the FA pathway could be involved in the coordination of both cell cycle checkpoints and DNA repair.
Fanconi anemia C protein acts at a switch between apoptosis and necrosis in mitomycin C-induced cell death
1999, Experimental Cell Research
Deregulation of apoptosis seems to be a hallmark of the Fanconi anemia (FA) syndrome. In order to further define the role of the FA protein from complementation group C (FAC) in apoptosis, we characterized parameters modified during the mitomycin-C (MMC)-induced apoptotic program. It is shown that despite a higher level of cell death for FA compared to normal lymphoblasts after MMC treatment, FA cells do not display a marked DNA fragmentation. Furthermore, while playing a central role in MMC apoptosis of normal lymphoblasts, the activity of caspase-3-like proteases is altered in FA cells. Interestingly, the disruption of the mitochondrial transmembrane potential (Δψ), an early event that can lead to apoptotic or to necrotic death, is accomplished similarly in FA and in normal cells. Finally, it is shown that the overexpressed FAC protein inhibited the apoptotic steps, with the exception of the decrease of the Δψ. Altogether, our results indicate that the FAC protein acts at a step preceding the activation of the caspases and after the modification of the Δψ, a decision point at which cells can be pushed toward either apoptosis or necrosis and which, consequently, regulates the balance between the two modes of cell death.
Is Fanconi anemia caused by a defect in the processing of DNA damage?
1998, Mutation Research - DNA Repair
Fanconi anemia (FA) is an autosomal genetic disease characterized by a complex array of developmental disorders, a high predisposition to bone marrow failure and to acute myelogenous leukemia. The chromosomal instability and the hypersensitivity to DNA cross-linking agents led to its classification with the DNA repair disorders. This review aimed at establishing whether it is still appropriate to consider 1\~FA within a DNA repair framework taking into account the recently discovered genetic heterogeneity characteristics of the defect (eight complementation groups). We discuss the possibility that the FA proteins interact to form a complex which may control different functions, including the processing of specific DNA lesions. Such a complex may act as a sensor to initiate protective systems as well as transcription of specific genes specifying, among others proteins, growth factors. Such steps may be organized as a linear cascade or more likely under the form of a web network.
Apoptosis and morphologic changes in drug-treated trabecular meshwork cells in vitro
1998, Experimental Eye Research
Using an in vitro culture system, we investigated whether bovine trabecular meshwork cells undergo apoptosis (programmed cell death) following exposure to anti-glaucoma medications (timolol, pilocarpine and epinephrine) and known inducers of apoptosis (5-fluorouracil, mitomycin-C and dexamethasone). Third to fifth passage bovine trabecular meshwork cells were grown to confluence and incubated for 1–12 days in growth media with timolol (1–1000 μm), pilocarpine (15–15000 μm), epinephrine (5–5000 μm), 5-fluorouracil (10–100 μg ml⁻¹), mitomycin-C (0.01–100 μg ml⁻¹) and dexamethasone (0.01–100 μm). The cultures were evaluated for apoptosis by phase-contrast microscopy, transmission electron microscopy and in situ apoptosis labeling. 5-Fluorouracil (10–100 μg ml⁻¹), mitomycin-C (0.1–100 μg ml⁻¹) and epinephrine (500–5000 μm) induced apoptosis in a dose and time-dependent manner. Timolol, pilocarpine, and dexamethasone-treated specimens did not show evidence of apoptosis at any of the concentrations tested. Trabecular meshwork cells incubated in timolol (100–1000 μm) developed cytoplasmic granules, and specimens treated with pilocarpine (15000 μm) developed cytoplasmic vacuoles. These granules and vacuoles have the appearance of secondary lysosomes. Dexamethasone-treated cells developed an increased number of mitochondria. This study suggests that the trabecular meshwork may undergo apoptosis following exposure to 5-fluorouracil, mitomycin-C and epinephrine. Timolol, pilocarpine and dexamethasone did not induce apoptosis. However, these drugs can incite characteristic morphologic changes in cultured trabecular meshwork cells.
Resistance to apoptosis in Fanconi's anaemia
1997, FEBS Letters
Fanconi's anaemia (FA) is a rare autosomal recessive disease characterised by progressive pancytopoenia, a diverse assortment of congenital malformations, an increased sensitivity to reactive oxygen species and a predisposition to the development of malignancies. In the present study, we assessed the propensity to undergo apoptosis of peripheral blood mononuclear cells (PBMC) from Italian FA patients. Cells were challenged by 2-deoxy-d-ribose (dRib) or TNF-α plus cycloheximide as agents that induce apoptosis by interfering with cell redox status and mitochondrial membrane potential (MMP), and PBMC from FA patients resulted to be less prone to die than those from healthy subjects. The decreased susceptibility of FA cells to undergo apoptosis was also evident when another parameter highly correlated with the apoptotic process, i.e. MMP, was measured. Moreover, when N-acetylcysteine was added to dRib-treated PBMC, a strong protection was evident either in PBMC from control subjects or from FA patients. These data indicate that an alteration of unknown nature of the mechanisms favouring apoptosis is present in freshly collected cells from FA patients, and that such alteration could contribute to the pathogenesis of the disease, and particularly to the increased susceptibility to cancer.
Distinct cellular phenotype linked to defective DNA interstrand crosslink repair and homologous recombination
2017, Molecular Medicine Reports

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^☆: This research was supported by grants from Krebsforschung Schweiz (No. AKT 322), from the Cancer League of both cantons of Basel, and from the Swiss National Fond (No. 32-9261.87).

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Short communicationApoptosis is not involved in the hypersensitivity of fanconi anemia cells to mitomycin C☆

Abstract

Biochem Pharmacol

Mutat Res

J Immunol Methods

Recessive inherited deficiencies predisposing to cancer

Anticancer Res

Fanconi anemia: constitutional aplastic anemia

Sem Hematol

Abnormal lymphokine production: a novel feature of the genetic disease Fanconi anemia

Hum Genet

Identification of two complementation groups in Fanconi anemia

Somatic Cell Molec Genet

Evidence for at least four Fanconi anaemia genes including FACC on chromosome 9

Nature Genet

Short communication
Apoptosis is not involved in the hypersensitivity of fanconi anemia cells to mitomycin C☆