In vitro Effects of Pentoxifylline and Doxorabicin on Cell Survival and DNA Damage in Sensitive and MDRP388 Leukemia Cells

It has been suggested that particle size plays an important role in determining the genotoxicity of gold nanoparticles (GNPs). The purpose of this study was to compare the potential radio-sensitization effects of two different sized GNPs (3.9 and 37.4 nm) fabricated and examined in vitro in Lewis Lung carcinoma (LLC) as a model of non-small cell lung cancer through use of comet and clonogenic assays. After the treatment of 2Gy X-ray irradiation, both particle sizes demonstrated increased DNA damage when compared to treatment with particles only and radiation alone. This radio-sensitization was further translated into a reduction in cell survival demonstrated by clonogenicity. This work indicates that GNPs of both sizes induce DNA damage in LLC cells at the tested concentrations, whereas the 37.4 nm particle size treatment group demonstrated greater significance in vitro. The presented data aids in the evaluation of the radiobiological response of Lewis Lung carcinoma cells treated with gold nanoparticles.

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In vitro drug sensitivity of cells from children with leukemia using the MTT assay with improved culture conditions

Blood ◽

10.1182/blood.v76.11.2327.2327 ◽

1990 ◽

Vol 76 (11) ◽

pp. 2327-2336 ◽

Cited By ~ 164

Author(s):

R Pieters ◽

AH Loonen ◽

DR Huismans ◽

GJ Broekema ◽

MW Dirven ◽

...

Keyword(s):

Drug Resistance ◽

Cell Survival ◽

Mtt Assay ◽

Large Scale ◽

Drug Sensitivity ◽

Lymphoblastic Leukemia ◽

Leukemia Cells ◽

Response Curves ◽

Mtt Reduction

Abstract The knowledge about drug resistance in childhood leukemias and acute lymphoblastic leukemia (ALL) in general is limited. This is because of the lack of a suitable in vitro drug sensitivity assay, which is in part due to low in vitro ALL cell survival. We recently adapted the highly efficient 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay to test cells from ALL patients and showed that its results were comparable with those of the DiSC assay, up to now the most valid but laborious assay. In this study, in vitro drug sensitivity was assessed in cells from 82 children with leukemia, 79 of whom had ALL, with the MTT assay. Dose response curves were obtained for 6-mercaptopurine, 6-thioguanine (6-TG), prednisolone (Pred), daunorubicin (DNR), vincristine (VCR), cytosine arabinoside (Ara-C), L- asparaginase (L-Asp), mafosfamide, and mustine. A cytotoxic effect of methotrexate could be detected in only a few cases. Large interindividual differences in drug sensitivity were detected. Compared with leukemia cells from newly diagnosed patients, leukemia cells from relapsed patients were significantly more in vitro resistant to 6-TG, Pred, Ara-C, mafosfamide and mustine but not to DNR, VCR, and L-Asp. Improvements of culture medium and methods to increase MTT reduction were studied. From 10 components tested, addition of insulin and bovine serum albumin to serum-containing medium improved ALL cell survival. Addition of succinate did not increase the amount of MTT reduction. We conclude that the in vitro MTT assay highly facilitates large-scale studies on drug resistance of ALL patients that can lead to rational improvements in existing treatment protocols.

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In vitro Effects of New Generation Oxazaphosphorines on Human Promyelocytic Leukemia Cells*

Folia Biologica ◽

10.3409/fb61_1-2.31 ◽

2013 ◽

Vol 61 (1) ◽

pp. 31-40 ◽

Cited By ~ 3

Author(s):

Lidia Mazur ◽

Małgorzata Opydo-Chanek ◽

Marta Stojak ◽

Ulf Niemeyer

Keyword(s):

Promyelocytic Leukemia ◽

Leukemia Cells ◽

In Vitro Effects ◽

New Generation

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IN VITRO EFFECTS OF COAL FLY ASHES: Hydroxyl Radical Generation, Iron Release, and DNA Damage and Toxicity in Rat Lung Epithelial Cells

Inhalation Toxicology ◽

10.1080/089583799196628 ◽

1999 ◽

Vol 11 (12) ◽

pp. 1123-1141 ◽

Cited By ~ 85

Author(s):

Jan M. S. van Maanen, Paul J. A. Bo

Keyword(s):

Dna Damage ◽

Lung Epithelial Cells ◽

Rat Lung ◽

Iron Release ◽

Fly Ashes ◽

Lung Epithelial ◽

Coal Fly Ashes ◽

Radical Generation ◽

In Vitro Effects

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DNA damage induced by phenylalanine and its analogue p-chlorophenylalanine in blood and brain of rats subjected to a model of hyperphenylalaninemia

Biochemistry and Cell Biology ◽

10.1139/bcb-2013-0023 ◽

2013 ◽

Vol 91 (5) ◽

pp. 319-324 ◽

Cited By ~ 11

Author(s):

Kellen R. Simon ◽

Rosane M. dos Santos ◽

Giselli Scaini ◽

Daniela D. Leffa ◽

Adriani P. Damiani ◽

...

Keyword(s):

Dna Damage ◽

Cerebral Cortex ◽

Phenylalanine Hydroxylase ◽

Neurological Dysfunction ◽

Control Group ◽

Acute Administration ◽

Dna Migration ◽

In Vitro Effects

Phenylketonuria (PKU) is a disease caused by a deficiency of phenylalanine hydroxylase (PAH), resulting in an accumulation of phenylalanine (Phe) in the brain tissue, cerebrospinal fluid, and other tissues of PKU patients. Considering that high levels of Phe are associated with neurological dysfunction and that the mechanisms underlying the neurotoxicity in PKU remain poorly understood, the main objective of this study was to investigate the in vivo and in vitro effects of Phe on DNA damage, as determined by the alkaline comet assay. The results showed that, compared to control group, the levels of DNA migration were significantly greater after acute administration of Phe, p-chlorophenylalanine (p-Cl-Phe, an inhibitor of PAH), or a combination thereof in cerebral cortex and blood, indicating DNA damage. These treatments also provoked increase of carbonyl content. Additionally, when Phe or p-Cl-Phe was present in the incubation medium, we observed an increase in the frequency and index of DNA damage in the cerebral cortex and blood, without affecting lactate dehydrogenase (LDH) release. Our in vitro and in vivo findings indicate that DNA damage occurs in the cerebral cortex and blood of rats receiving Phe, suggesting that this mechanism could be, at least in part, responsible for the neurological dysfunction in PKU patients.

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Lenalidomide Abrogates the Protective Influence of Nurse-Like Cells on Primary Chronic Lymphocytic Leukemia Cells In Vitro.

Blood ◽

10.1182/blood.v110.11.3116.3116 ◽

2007 ◽

Vol 110 (11) ◽

pp. 3116-3116 ◽

Cited By ~ 2

Author(s):

Danelle F. James ◽

Maryann R. Betty ◽

Ruzbeh Mosadeghi ◽

Thomas J. Kipps

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Cell Viability ◽

Cell Survival ◽

Leukemia Cell ◽

Lymphocytic Leukemia ◽

Response To Treatment ◽

Leukemia Cells ◽

High Dependency ◽

Cells Cultured

Abstract Lenalidomide (3-(4-amino-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione)) is an agent approved for treatment of patients with del 5q myelodysplastic syndromes and previously treated multiple myeloma. Lenalidomide has been found in early clinical trials to have potential therapeutic activity in patients with relapsed chronic lymphocytic leukemia (CLL). The mechanism(s) whereby this drug is active in CLL is unknown. In particular, studies to date have not found lenalidomide to have any direct cytotoxic activity on CLL cells in vitro. This has stimulated speculation that this agent might adversely affect the positive influence of the microenvironment on leukemia-cell survival. We and others have observed that cells found in the leukemia microenvironment can support CLL-cell survival in vitro. One such type of cells are nurse-like cells (NLC), which can differentiate from the CD14-positive blood mononuclear cells of CLL patients into large, round adherent cells that can attract and support CLL cell survival in vitro for weeks, if not longer. We evaluated the effects of lenalidomide on primary leukemia-cell survival in vitro when the CLL cells from different patients (N=21) were cultured alone or together with NLC generated as previously described [Tsukada Blood 2002]. We assessed the in-vitro activity of lenalidomide on primary CLL cells from 21 patients, in duplicate in a series of 6 experiments. Lenalidomide at concentrations of 0.1μM-200μM did not significantly impact the survival of CLL cells that were cultured alone for up to 12 days. Analysis of cell surface markers revealed increased expression of CD38 at 36 hours in 5/5 lenalidomide treated CLL samples compared with untreated cells (MFIR 5.7 +/− .86 vs. 3.4 +/− .83 p=.003). We observed sustained upregualtion of CD40 and regulation of CXCR4 in the majority of cells treated with lenalidomide. When cultured with NLC, the survival of CLL cells was comparable to or significantly higher than that of CLL cells cultured alone 62.4% vs. 51% (+/−3% SEM n=21 p [<] 0.0005). The addition of lenalidomide at concentrations of 0.1μM and greater to co-cultures of NLC and CLL cells caused specific reductions in CLL cell survival to levels similar to or lower than that of CLL cells cultured without NLC. In the presence of NLC, lenalidomide at 1μM reduced CLL cell viability compared to control (41.5% vs. 56% +/−4% p [<] 0.0005 paired student t test n=13). For most patients the levels of CLL cell viability on days 4 through 8 in the co-cultures with lenalidomide was significantly lower than those of CLL cells co-cultured with NLC in the absence of lenalidomide. As such, this study reveals that physiologic concentrations of lenalidomide might abrogate the protective influence of NLC on CLL cell survival in vitro and potentially in vivo. Conceivably, those patients who have leukemia cells displaying a high dependency on NLC for survival in vitro also might be most likely to experience a favorable clinical response to treatment with lenalidomide. This hypothesis will be tested in a prospective manner with a planned clinical trial evaluating lenalidomide for treatment of CLL through the CLL Research Consortium.

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