Identification of transforming growth factor-?1-binding protein overexpression in carmustine-resistant glioma cells by MRNA differential display

Norman, Sylvia A.; Treasurywala, Sherri; Hoelzinger, Dominique B.; Shapiro, Joan Rankin; Scheck, Adrienne C.

doi:10.1002/1097-0142(20000815)89:4<850::aid-cncr18>3.0.co;2-b

Cited by 8 publications

(3 citation statements)

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“…In a study using mRNA differential display, BCNU has been shown to increase the expression of a gene encoding the latent transforming growth factor-binding protein 1 (LTBP-1) by three-fold suggesting that BCNU might increase transforming growth factor-β1 (TGF-β1) signaling [35], which is known to play a pivotal role in APP metabolism. Therefore, to understand the possible molecular mechanism for the reduced amyloidogenic processing of APP by BCNU, we quantified the levels of TGFβ protein released into the CM and also in the lysates after cells were treated with different concentrations of BCNU.…”

Section: Resultsmentioning

confidence: 99%

Striking reduction of amyloid plaque burden in an Alzheimer's mouse model after chronic administration of carmustine

Hayes

Dey

Palavicini

et al. 2013

BMC Med

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BackgroundCurrently available therapies for Alzheimer's disease (AD) do not treat the underlying cause of AD. Anecdotal observations in nursing homes from multiple studies strongly suggest an inverse relationship between cancer and AD. Therefore, we reasoned that oncology drugs may be effective against AD.MethodsWe screened a library of all the FDA-approved oncology drugs and identified bis-chloroethylnitrosourea (BCNU or carmustine) as an effective amyloid beta (Aβ) reducing compound. To quantify Aβ levels, Chinese hamster ovary (CHO) cells stably expressing amyloid precursor protein 751WT (APP751WT) called 7WD10 cells were exposed to different concentrations of BCNU for 48 hours and the conditioned media were collected. To detect Aβ the conditioned media were immunoprecipitated with Ab9 antibody and subjected to immunoblot detection. Amyloid plaques were quantified in the brains of a mouse model of AD after chronic exposure to BCNU by thoflavin S staining.ResultsBCNU decreased normalized levels of Aβ starting from 5 μM by 39% (P < 0.05), 10 μM by 51% (P < 0.01) and 20 μM by 63% (P < 0.01) in CHO cells compared to a control group treated with butyl amine, a structural derivative of BCNU. Interestingly, soluble amyloid precursor protein α (sAPPα) levels were increased to 167% (P < 0.01) at 0.5 μM, 186% (P < 0.05) at 1 μM, 204% (P < 0.01) at 5 μM and 152% (P < 0.05) at 10 μM compared to untreated cells. We also tested the effects of 12 structural derivatives of BCNU on Aβ levels, but none of them were as potent as BCNU. BCNU treatment at 5 μM led to an accumulation of immature APP at the cell surface resulting in an increased ratio of surface to total APP by 184% for immature APP, but no change in mature APP. It is also remarkable that BCNU reduced Aβ generation independent of secretases which were not altered up to 40 μM. Interestingly, levels of transforming growth factor beta (TGFβ) were increased at 5 μM (43%, P < 0.05), 10 μM (73%, P < 0.01) and 20 μM (92%, P < 0.001). Most significantly, cell culture results were confirmed in vivo after chronic administration of BCNU at 0.5 mg/kg which led to the reduction of Aβ40 by 75% and amyloid plaque burden by 81%. Conversely, the levels of sAPPα were increased by 45%.ConclusionsBCNU reduces Aβ generation and plaque burden at non-toxic concentrations possibly through altered intracellular trafficking and processing of APP. Taken together these data provided unequivocal evidence that BCNU is a potent secretase-sparing anti-Aβ drug.See related commentary article here http://www.biomedcentral.com/1741-7015/11/82

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Section: Resultsmentioning

confidence: 99%

Striking reduction of amyloid plaque burden in an Alzheimer's mouse model after chronic administration of carmustine

Hayes

Dey

Palavicini

et al. 2013

BMC Med

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“…The passage number of the cells used for all experiments varied (LX/LXR 15–32, DI 5–25, DIR 15–40, ME 18–41; MER 52–120); however, the results of repeats done at different passage numbers were consistent. Cells were selected for resistance to 10 μg/ml BCNU as described [ 10 , 12 ], and mock treatment of the cells was done in parallel. Briefly, cells were washed with MAB without serum 3 times; they were then mock-treated using MAB without serum or drug, or treated with BCNU in MAB without serum for 1 hour at 37°C with 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Over-representation of specific regions of chromosome 22 in cells from human glioma correlate with resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea

2006

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BackgroundGlioblastoma multiforme is the most malignant form of brain tumor. Despite treatment including surgical resection, adjuvant chemotherapy, and radiation, these tumors typically recur. The recurrent tumor is often resistant to further therapy with the same agent, suggesting that the surviving cells that repopulate the tumor mass have an intrinsic genetic advantage. We previously demonstrated that cells selected for resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) are near-diploid, with over-representation of part or all of chromosomes 7 and 22. While cells from untreated gliomas often have over-representation of chromosome 7, chromosome 22 is typically under-represented.MethodsWe have analyzed cells from primary and recurrent tumors from the same patient before and after in vitro selection for resistance to clinically relevant doses of BCNU. Karyotypic analyses were done to demonstrate the genetic makeup of these cells, and fluorescent in situ hybridization analyses have defined the region(s) of chromosome 22 retained in these BCNU-resistant cells.ResultsKaryotypic analyses demonstrated that cells selected for BCNU resistance were near-diploid with over-representation of chromosomes 7 and 22. In cells where whole copies of chromosome 22 were not identified, numerous fragments of this chromosome were retained and inserted into several marker and derivative chromosomes. Fluorescent in situ hybridization analyses using whole chromosome paints confirmed this finding. Additional FISH analysis using bacterial artificial chromosome probes spanning the length of chromosome 22 have allowed us to map the over-represented region to 22q12.3–13.32.ConclusionCells selected for BCNU resistance either in vivo or in vitro retain sequences mapped to chromosome 22. The specific over-representation of sequences mapped to 22q12.3–13.32 suggest the presence of a DNA sequence important to BCNU survival and/or resistance located in this region of chromosome 22.

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“…Many studies have shown the differential expression changes of drug resistance-related genes in glioma cells aquiring drug resistance [3]. However, there have been few reports about gene expression changes beside drug resistance-related genes [4]. In Japan, 1-(4-amino-2-methyl-5-pyrimidinyl)-methyl-3-(2-chloroethyl)-3-nitrosourea (nimustine hydrocloride, ACNU) is the most common chemotherapeutic agent used in glioma therapy [5,6].…”

mentioning

confidence: 99%

Gene expression profiles of 1-(4-amino-2-methyl-5-pyrimidinyl)-methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU)-resistant C6 rat glioma cells

et al. 2006

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Chemotherapy in itself is suspected to cause the development or selection of drug-resistant tumor cells, which have more aggressive phenotypes. The authors investigated the differential changes of gene expression in the 1-(4-amino-2-methyl-5-pyrimidinyl)-methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU)-resistant subline of the C6 rat glioma (C6AR2), which was established from C6 rat glioma cells by exposure to ACNU in vitro. The resistance to ACNU of C6AR2 was confirmed by MTS assay. The increased expression of O6-methylguanine-DNA methyltransferase in C6AR2 cells was shown using RT-PCR. C6AR2 cells displayed a higher proliferative activity relative to C6 cells. Analysis with cDNA array showed that 19 genes were transcriptionally up-regulated and 16 genes down-regulated in C6AR2 cells compared to C6 cells. They belonged to various functional classes of genes beside the drug-resistant system. Among them, the down-regulation of several genes in C6AR2 cells, including c-kit, pleiotrophin, platelet-derived growth factor receptor-alpha, peripheral myelin protein-22 and NG2 chondroitin sulfate proteoglycan, which are expressed originally in developmental glial lineages, were verified using semi-quantitative RT-PCR. In addition, the gene expression of astroglial intermediate filament proteins, including GFAP, vimentin and nestin, were decreased in C6AR2 cells relative to C6 cells in semi-quantitative RT-PCR and immunocytochemistry. These findings may represent an undifferentiated state of ACNU-resistant glioma cells and a more aggressive phenotype in recurrent tumors following chemotherapy.

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Identification of transforming growth factor-?1-binding protein overexpression in carmustine-resistant glioma cells by MRNA differential display

Cited by 8 publications

References 42 publications

Striking reduction of amyloid plaque burden in an Alzheimer's mouse model after chronic administration of carmustine

Striking reduction of amyloid plaque burden in an Alzheimer's mouse model after chronic administration of carmustine

Over-representation of specific regions of chromosome 22 in cells from human glioma correlate with resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea

Gene expression profiles of 1-(4-amino-2-methyl-5-pyrimidinyl)-methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU)-resistant C6 rat glioma cells

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