Methylation of human ornithine decarboxylase gene before transfection abolishes its transient expression in Chinese hamster ovary cells

Halmekytö, Maria; Hirvonen, Ari; Wahlfors, Jarmo; Alhonen, Leena; Jänne, Juhani

doi:10.1016/0006-291x(89)92029-9

Cited by 16 publications

(9 citation statements)

References 17 publications

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“…DNA methylation is established as a mechanism regulating ODC1 gene expression. In early studies a positive correlation between ODC1 gene hypomethylation and expression was reported 40 and experimental methylation of the ODC1 gene abolished its expression 41 . Furthermore aberrant ODC1 methylation has been reported in malignant cells 42 .…”

Section: Discussionmentioning

confidence: 98%

Basic Hallmarks of Urothelial Cancer Unleashed in Primary Uroepithelium by Interference with the Epigenetic Master Regulator ODC1

Erichsen

Seifert

Schulz

et al. 2020

Sci Rep

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Urothelial carcinoma (UC) is a common disease causing significant morbidity and mortality as well as considerable costs for health systems. extensive aberrant methylation of DnA is broadly documented in early Uc, contributing to genetic instability, altered gene expression and tumor progression. However the triggers initiating aberrant methylation are unknown. Recently we discovered that several genes encoding key enzymes of methyl group and polyamine metabolism, including Ornithine Decarboxylase 1 (ODC1), are affected by DNA methylation in early stage UC. In this study, we investigated the hypothesis that these epigenetic alterations act in a feed-forward fashion to promote aberrant DnA methylation in Uc. We demonstrate that siRnA-mediated knockdown of ODC1 expression elicits genome-wide LINE-1 demethylation, induction of LINE-1 transcripts and doublestrand DnA breaks and decreases viability in primary cultured uroepithelial cells. Similarly, following siRnA-mediated knockdown of ODC1, Uc cells undergo double-strand DnA breaks and apoptosis. Collectively, our findings provide evidence that ODC1 gene hypermethylation could be a starting point for the onset of genome-wide epigenetic aberrations in urothelial carcinogenesis. Furthermore, LINE-1 induction enabled by ODC1 interference provides a new experimental model to study mechanisms and consequences of LINE-1 activation in the etiology and progression of UC as well as presumably other cancers. Urothelial carcinoma (UC), the most common cancer of the urinary bladder, is a frequent disease with yearly 549,393 new cases and 199,922 deaths worldwide 1. Relapse rates of 30-70% and rates of progression at 10-30% for high-grade tumors present a serious health care challenge and impose enormous financial burdens on the health care systems 2. In particular, UC that have invaded the muscle-layers of the bladder are highly lethal. Tobacco smoking is thought to contribute to 50% of all UC cases as a risk factor 3. Aromatic amines such as 2-naphthylamin and polycyclic aromatic hydrocarbons in cigarette smoke cause mutations in key cancer-related genes by forming DNA adducts 4. Accordingly, exposure to tobacco smoke alone is sufficient to induce

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Section: Discussionmentioning

confidence: 98%

Basic Hallmarks of Urothelial Cancer Unleashed in Primary Uroepithelium by Interference with the Epigenetic Master Regulator ODC1

Erichsen

Seifert

Schulz

et al. 2020

Sci Rep

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“…However, upon serum stimulation of the longterm quiescent WI-38 cells, steady-state levels of ODC mRNA begin to increase between 5 and 7 h following stimulation and reach maximum levels by 10 h after stimulation . ODC expression has been shown to be regulated at the transcriptional level in human diploid fibroblasts stimulated with mitogens (Abrahamsen and Morris, 1990;Brabant et al, 1988;Halmekyto et al, 1989;Holtta et al, 1989;Kahana and Nathans, 1984;Katz and Kahana, 1987;Lu et al, 1991;Manzella and Blackshear, 1990;Manzella et al, 1991;Olsen and Spizz, 1986;Palvimo et al, 1991;Van Daalen Wetters et al, 1989a,b). It was likely, therefore that the differences previously observed in steady-state levels of ODC mRNA in short-and long-term cells were due, at least in part to differences at the transcriptional level.…”

Section: Odc Is Transcriptionally Regulated In Wi-38 Cells Stimulatedmentioning

confidence: 99%

“…ODC levels have been shown to be highly regulated in response to a large number of stimuli affecting cell growth and differentiation, including growth factors, steroid hormones, phorbol esters, and CAMP elevating agents (Manzella et al, 1991;Olsen and Spizz, 1986;Pegg, 1986Pegg, , 1988Tabor and Tabor, 1984;Van Daalen Wetters et al, 1989a,b). This regulation has been shown to occur a t multiple levels, including changes in the rate of transcription, the stability and translational efficiency of the ODC mRNA, and stability of the enzyme protein (Abrahamsen and Morris, 1990;Brabant et al, 1988;Halmekyto et al, 1989;Holtta et al, 1989;Kahana and Nathans, 1984;Katz and Kahana, 1987;Lu et al, 1991;Manzella and Blackshear, 1990;Manzella et al, 1991;Olsen and Spizz, 1986;Palvimo et al, 1991;Van Daalen Wetters et al, 1989a,b).…”

Section: Incmentioning

confidence: 99%

Regulation of human ornithine decarboxylase expression following prolonged quiescence: Role for the c‐Myc/Max protein complex

Peña

Hickok

et al. 1995

Journal Cellular Physiology

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WI-38 cells can remain quiescent for long periods of time and still be induced to reenter the cell cycle by the addition of fresh serum. However, the longer these cells remain growth arrested, the more time they require to enter S phase. This prolongation of the prereplicative phase has been localized to a point early in G1, after the induction of "immediate early" G1 genes such as c-fos and c-jun but before maximal expression of "early" G1 genes such as ornithine decarboxylase (ODC). Understanding the molecular basis for ODC mRNA induction can therefore provide information about the molecular events which regulate the progression of cells out of long-term quiescence into G1 and subsequently into DNA synthesis. Studies utilizing electrophoretic mobility shift assays (EMSA) of nuclear extracts from short- and long-term quiescent WI-38 cells identified a region of the human ODC promoter at -491 bp to -474 bp which exhibited a protein binding pattern that correlated with the temporal pattern of ODC mRNA expression. The presence of a CACGTG element within this fragment, studies with antibodies against c-Myc and Max, the use of purified recombinant c-Myc protein in the mobility shift assay, and antisense studies suggest that these proteins can specifically bind this portion of the human ODC promoter in a manner consistent with growth-associated modulation of the expression of ODC and other early G1 genes following prolonged quiescence. These studies suggest a role for the c-Myc/Max protein complex in regulating events involved in the progression of cells out of long-term quiescence into G1 and subsequently into S.

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“…Transgenic mice were produced using the standard microinjection technique (Hogan et al, 1986). The transgene construct was a cloned human ODC gene previously shown to be expressed actively both persistently (Holtta et al, 1989) and transiently (Halmekyto et al, 1989) in transfected cells. The construct contained, in addition to the I2 exons and 1 1 introns of the gene (Hickok et al, 1990), some 800 nucleotides of the 5' flanking and 1,000 nucleotides of the 3' flanking regions.…”

Section: Animalsmentioning

confidence: 99%

Nuclear Magnetic Resonance Spectroscopy Study on Energy Metabolism, Intracellular pH, and Free Mg²⁺ Concentration in the Brain of Transgenic Mice Overexpressing Human Ornithine Decarboxylase Gene

Kauppinen

Halmekytö

Alhonen

et al. 1992

Journal of Neurochemistry

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We have generated a transgenic mouse line strikingly overexpressing the human ornithine decarboxylase (ODC) gene in their brain. Brain ODC activity was increased in the transgenic animals by a factor of 70 in comparison with their nontransgenic littermates. The content of brain putrescine, the product of ODC, was greater than 60 mumol/g of tissue in the transgenic mice, whereas in the normal animals it was below the level that could be detected by an HPLC method. The concentrations of the higher polyamines (spermidine and spermine) were not significantly different from control values. 31P nuclear magnetic resonance (31P NMR) spectroscopy analyses revealed a significantly reduced (40%) free Mg2+ concentration as calculated from the chemical shift differences of the nucleoside triphosphate alpha and beta peaks in the brains of the transgenic animals. The lower free Mg2+ concentration in the brains of ODC transgenic mice was not a consequence of altered intracellular pH or changes in cellular high-energy metabolites. 1H NMR showed no differences in brain choline/N-acetylaspartate and total creatine/N-acetylaspartate ratios between the two animal groups. These ODC transgenic animals may serve as models in vivo for studies on cerebral postischemic events and on epilepsy, as polyamines are supposed to be involved in these processes.

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Methylation of human ornithine decarboxylase gene before transfection abolishes its transient expression in Chinese hamster ovary cells

Cited by 16 publications

References 17 publications

Basic Hallmarks of Urothelial Cancer Unleashed in Primary Uroepithelium by Interference with the Epigenetic Master Regulator ODC1

Basic Hallmarks of Urothelial Cancer Unleashed in Primary Uroepithelium by Interference with the Epigenetic Master Regulator ODC1

Regulation of human ornithine decarboxylase expression following prolonged quiescence: Role for the c‐Myc/Max protein complex

Nuclear Magnetic Resonance Spectroscopy Study on Energy Metabolism, Intracellular pH, and Free Mg²⁺ Concentration in the Brain of Transgenic Mice Overexpressing Human Ornithine Decarboxylase Gene

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Methylation of human ornithine decarboxylase gene before transfection abolishes its transient expression in Chinese hamster ovary cells

Cited by 16 publications

References 17 publications

Basic Hallmarks of Urothelial Cancer Unleashed in Primary Uroepithelium by Interference with the Epigenetic Master Regulator ODC1

Basic Hallmarks of Urothelial Cancer Unleashed in Primary Uroepithelium by Interference with the Epigenetic Master Regulator ODC1

Regulation of human ornithine decarboxylase expression following prolonged quiescence: Role for the c‐Myc/Max protein complex

Nuclear Magnetic Resonance Spectroscopy Study on Energy Metabolism, Intracellular pH, and Free Mg2+ Concentration in the Brain of Transgenic Mice Overexpressing Human Ornithine Decarboxylase Gene

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Nuclear Magnetic Resonance Spectroscopy Study on Energy Metabolism, Intracellular pH, and Free Mg²⁺ Concentration in the Brain of Transgenic Mice Overexpressing Human Ornithine Decarboxylase Gene