Comparison of the in vitro cytotoxicity (L1210) of 5-Aza-2′-deoxycytidine with its therapeutic and toxic effects in mice

Covey, Joseph M.; Zaharko, Daniel S.

doi:10.1016/0277-5379(85)90207-x

Cited by 25 publications

(22 citation statements)

References 4 publications

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“…Many methylationmodifying chemical agents, notably the nucleoside analogues 5-azacytidine or 5-aza-deoxycytidine, are known to be capable of activating the expression of a number of tumor-specific, methylation-repressed genes in cancer cells [24][25][26] . However, their clinical use is limited by their cytotoxicity and nonspecific effects on both normal and cancer cells [27] . There is a need to develop novel, specific methylation-modifying agents with minimal toxicity.…”

Section: Discussionmentioning

confidence: 99%

Quercetin Is Able to Demethylate the p16INK4a Gene Promoter

Tan

Wang²,

Lu³

et al. 2008

Chemotherapy

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Background: Quercetin is a flavonoid found ubiquitously in nature. Studies in vitroand in vivohave suggested that quercetin may have a protective role against colon cancer. Methods: We selected the human colon cancer cell line RKO to investigate the effects of quercetin in vitro. RKO cells were treated with different concentrations of quercetin. Results: In comparison to the control, quercetin was able to inhibit the growth of RKO cells, as measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Untreated RKO cells demonstrated almost complete methylation of the p16INK4a gene. Hypermethylation of the p16INK4a gene was successfully reversed after 120 h of treatment with quercetin. Expression of the p16INK4a gene was restored in a concentration-dependent manner. Conclusion: All these data suggest that quercetin has antitumor properties, probably via demethylation of the p16INK4a gene promoter.

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Section: Discussionmentioning

confidence: 99%

Quercetin Is Able to Demethylate the p16INK4a Gene Promoter

Tan

Wang²,

Lu³

et al. 2008

Chemotherapy

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“…A second is a highperformance liquid chromatography/ultraviolet (HPLC-UV) method and has a LLOQ of 50 ng/mL in human plasma. 36,37 Both of these methods suffer from low sensitivity and may not be specific. Additionally, although both of these methods were used to characterize decitabine pharmacokinetics at high doses (e.g.…”

Section: -Aza-2mentioning

confidence: 99%

“…Additionally, although both of these methods were used to characterize decitabine pharmacokinetics at high doses (e.g. 2 mg/kg in monkeys and 70-100 mg/m 2 in patients 37,38 ), none could be applied at lower doses. In fact, no low dose pharmacokinetics of decitabine has been published.…”

Section: -Aza-2mentioning

confidence: 99%

Characterization of decomposition products and preclinical and low dose clinical pharmacokinetics of decitabine (5‐aza‐2′‐deoxycytidine) by a new liquid chromatography/tandem mass spectrometry quantification method

Liu

Marcucci

Byrd

et al. 2006

Rapid Comm Mass Spectrometry

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Aberrant DNA methylation patterns resulting in gene transcriptional repression are observed in numerous cancers. Decitabine, a DNA methyltransferase inhibitor, is being clinically evaluated in patients with hematologic malignancies and solid tumors. Decitabine is rather unstable and decomposes to 1-beta-D-2'-deoxyribofuranosyl-3-guanylurea under basic conditions and several additional unknown products under neutral conditions. This has greatly limited application of pharmacokinetic assays to clinical development of decitabine. In this paper, a high-performance liquid chromatography/ultraviolet multi-stage mass spectrometry (HPLC-UV-MSn) study of the decomposition of decitabine in water and human plasma revealed that these previously unknown products are isomers of the intermediates formyl-1-beta-D-2'-deoxyribofuranosyl-3-guanylurea and 1-beta-D-2'-deoxyribofuranosyl-3-guanylurea. A HPLC tandem mass spectrometry (MS/MS) method for the determination of decitabine concentrations in human and rat plasma has been developed. This method was based on a specific fragmentation pathway of the molecular ion of decitabine at m/z 229 to generate a unique fragment ion at m/z 113 under collision-induced dissociation. Separation of decitabine and the stable internal standard dihydro-5-aza-cytidine from the endogenous interfering substance in plasma extract was carried out on a C18 Aquasil column under an isocratic elution with a mobile phase consisting of 5% water/acetonitrile and 10 mM ammonium formate. The detection of decitabine was via selected reaction monitoring (SRM, 229 > 113), and its ionization was enhanced by post-column addition of acetonitrile. Effects of sample preparation and handling parameters on the stability of decitabine were also evaluated in human plasma at various temperatures. The accuracy and precision of this assay showed a coefficient of variation of <15% over the range of 0.5-25 ng for rat plasma and 0.1-25 ng for human plasma injected on-column. Pharmacokinetics of decitabine in rats following intravenous doses of 1.0 and 5.0 mg/kg were characterized. In the rat, plasma concentration-time profiles were found to follow a biexponential decline and the pharmacokinetics was dose-independent. Application of this decitabine pharmacokinetic assay to human studies is therefore justified and ongoing.

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“…After small tumors had grown, the mice were treated with 5-aza-dCyd as described and the tumor size and weight at sacrifice were measured. We based our 5-aza-dCyd dosing in the mice on what had been used in previous studies and chose the lowest doses that had been shown to be effective (52)(53)(54). We picked a low dose to minimize the cytotoxic effects of 5-aza-dCyd on tumor growth.…”

Section: Discussionmentioning

confidence: 99%

The In vitro and In vivo Effects of Re-Expressing Methylated von Hippel-Lindau Tumor Suppressor Gene in Clear Cell Renal Carcinoma with 5-Aza-2′-deoxycytidine

Alleman

Tabios

Chandramouli

et al. 2004

Clinical Cancer Research

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Purpose: Clear cell renal carcinoma (ccRCC) is strongly associated with loss of the von Hippel-Lindau (VHL) tumor suppressor gene. The VHL gene is functionally lost through hypermethylation in up to 19% of sporadic ccRCC cases. We theorized that re-expressing VHL silenced by methylation in ccRCC cells, using a hypo-methylating agent, may be an approach to treatment in patients with this type of cancer. We test the ability of two hypo-methylating agents to re-express VHL in cell culture and in mice bearing human ccRCC and evaluate the effects of re-expressed VHL in these models.Experimental Design: Real-time reverse transcription-PCR was used to evaluate the ability of zebularine and 5-aza-2-deoxycytidine (5-aza-dCyd) to re-express VHL in four ccRCC cell lines with documented VHL gene silencing through hypermethylation. We evaluated if the VHL reexpressed after hypo-methylating agent treatment could recreate similar phenotypic changes in ccRCC cells observed when the VHL gene is re-expressed via transfection in cell culture and in a xenograft mouse model. Finally we evaluate global gene expression changes occurring in our cells, using microarray analysis.Results: 5-Aza-dCyd was able to re-express VHL in our cell lines both in culture and in xenografted murine tumors.Well described phenotypic changes of VHL expression including decreased invasiveness into Matrigel, and decreased vascular endothelial growth factor and glucose transporter-1 expression were observed in the treated lines. VHL methylated ccRCC xenografted tumors were significantly reduced in size in mice treated with 5-aza-dCyd. Mice bearing nonmethylated but VHL-mutated tumors showed no tumor shrinkage with 5-aza-dCyd treatment.Conclusion: Hypo-methylating agents may be useful in the treatment of patients having ccRCC tumors consisting of cells with methylated VHL.

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Comparison of the in vitro cytotoxicity (L1210) of 5-Aza-2′-deoxycytidine with its therapeutic and toxic effects in mice

Cited by 25 publications

References 4 publications

Quercetin Is Able to Demethylate the p16INK4a Gene Promoter

Quercetin Is Able to Demethylate the p16INK4a Gene Promoter

Characterization of decomposition products and preclinical and low dose clinical pharmacokinetics of decitabine (5‐aza‐2′‐deoxycytidine) by a new liquid chromatography/tandem mass spectrometry quantification method

The In vitro and In vivo Effects of Re-Expressing Methylated von Hippel-Lindau Tumor Suppressor Gene in Clear Cell Renal Carcinoma with 5-Aza-2′-deoxycytidine

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