Prolonged culture of normal chorionic villus cells yields ICF syndrome-like chromatin decondensation and rearrangements

Tsien, Fern; Fiala, Emerich S.; Youn, Byungwoo; Long, Theodore; Laird, Peter W.; Weissbecker, Karen; Ehrlich, Melanie

doi:10.1159/000068543

Cited by 26 publications

(29 citation statements)

References 38 publications

(62 reference statements)

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“…We found that 3.3% (3.2770.17) of the cytosines were methylated in these tissues, which is virtually identical to values obtained in normal placenta (3.2270.08). In addition, these values are similar to the percentage of 5-methylcytosine determined by highperformance liquid chromatography in placenta (3.2%) and chorionic villi (3.1%) (Gama-Sosa et al, 1983a;Tsien et al, 2002). Moreover, identical levels of DNA methylation between molar tissues and placenta have been also observed at some single-copy sequences (Hjelle et al, 1982).…”

supporting

confidence: 82%

supporting

confidence: 68%

“…The lowest level found is in placenta, where 3.2% of the cytosines are methylated, and the highest level (4.26%) is found in brain (Tsien et al, 2002). As expected, LINE-1, Sata and Sat2 repeated sequences contain small amounts of methylated CpG in placenta compared to post-natal tissues (Gama-Sosa et al, 1983b;Tsien et al, 2002). However, base composition analysis indicates that Alu sequences are not hypomethylated in placenta and exhibit the same 5-methylcytosine content as brain Alu sequences (Gama-Sosa et al, 1983b).…”

supporting

confidence: 55%

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Specific hypermethylation of LINE-1 elements during abnormal overgrowth and differentiation of human placenta

Ballestar

Fraga

et al. 2006

Oncogene

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In human post-natal somatic cells, low global levels of DNA methylation have been associated with the hypomethylation of several repetitive elements, a feature that has been proposed to be a surrogate epigenetic marker. These data, mainly derived from the analysis of cancer cells, suggest a potential association between loss of cellgrowth control and altered differentiation with hypomethylation of repetitive sequences. Partial hydatidiform moles (PHMs) can be used as an alternative model for investigating this association in a non-tumorigenic context. This gestational disease is characterized by abnormal overgrowth and differentiation of the placenta and spontaneous abortion. Here, we comprehensively analyse the DNA methylation of these trophoblastic tissues in both PHM and normal placenta at global and sequencespecific levels. Analysis of the global 5-methylcytosine content and immunohistochemistry indicate that PHM and normal placenta have identical global levels of DNA methylation. In contrast, bisulfite genomic sequencing shows that, whereas Alu, NBL2 and satellite 2 repetitive elements are equally methylated, LINE-1 sequences are hypermethylated in PHM tissues (B2-fold relative to normal placenta). Interestingly, altered demethylation is also found in triploid diandric embryos that originate from dispermic fertilization of an oocyte, a common event responsible for most PHMs. In conclusion, alterations of DNA methylation do not seem to be randomly distributed in PHM, as several repeated elements remain unaltered, whereas LINE-1 sequences are hypermethylated. In addition, our findings suggest that the hypomethylation of repetitive elements in cancer is directly linked to the neoplasic process and not a simple consequence of loss of growth control observed in most of the cancer cells.

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supporting

confidence: 82%

supporting

confidence: 68%

supporting

confidence: 55%

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Specific hypermethylation of LINE-1 elements during abnormal overgrowth and differentiation of human placenta

Ballestar

Fraga

et al. 2006

Oncogene

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“…We have utilized the present procedure for chromosome isolation from cells of various organisms, including various cell types obtained from human, Rhesus macaques, rats, and mice 11,[20][21][22] . The standard protocol is provided, but certain key steps and variables may need to be adjusted for these diverse types of cells.…”

Section: Discussionmentioning

confidence: 99%

Chromosome Preparation From Cultured Cells

Howe

Umrigar

Tsien

2014

JoVE

106

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Chromosome (cytogenetic) analysis is widely used for the detection of chromosome instability. When followed by G-banding and molecular techniques such as fluorescence in situ hybridization (FISH), this assay has the powerful ability to analyze individual cells for aberrations that involve gains or losses of portions of the genome and rearrangements involving one or more chromosomes. In humans, chromosome abnormalities occur in approximately 1 per 160 live births 1,2 , 60-80% of all miscarriages 3,4

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“…Total demethylation of DNA was established in senescent cells (Tsien et al, 2002). Demethylation and decondensation of chromosome 1 pericentromeric satellites 2 and 3 in senescent fibroblasts were demonstrated by Suzuki et al (2002).…”

Section: Satdna Methylation and Degree Of Condensationmentioning

confidence: 99%

Human chromosome 1 satellite 3 DNA is decondensed, demethylated and transcribed in senescent cells and in A431 epithelial carcinoma cells

Enukashvily

Donev²,

Waisertreiger³

et al. 2007

Cytogenet Genome Res

103

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Constitutive heterochromatin mainly consists of different classes of satellite DNAs and is defined as a transcriptionally inactive chromatin that remains compact throughout the cell cycle. The aim of this work was to investigate the level of condensation, methylation and transcriptional status of centromeric (alphoid DNA) and pericentromeric satellites (human satellite 3, HS3) in tissues (lymphocytes, placenta cells) and in cultured primary (MRC5, VH-10, AT2Sp) and malignant (A431) cells. We found that alphoid DNA remained condensed and heavily methylated in all the cell types. The HS3 of chromosome 1 (HS3-1) but not of chromosome 9 (HS3-9) was strongly decondensed and demethylated in A431 cells. The same observation was made for aged embryonic lung (MRC5) and juvenile foreskin (VH-10) fibroblasts obtained at late passages (32^nd and 23^rd, respectively). Decondensation was also found in ataxia telangiectasia AT2Sp fibroblasts at the 16^th passage. One of the manifestations of the disease is premature aging. The level of HS3-1 decondensation was higher in aged primary fibroblasts as compared to A431. The HS3-1 extended into the territory of neighbouring chromosomes. An RT-PCR product was detected in A431 and senescent MRC5 fibroblasts using primers specific for HS3-1. The RNA was polyadenylated and transcribed from the reverse chain. Our results demonstrate the involvement of satellite DNA in associations between human chromosomes and intermingling of chromosome territories. The invading satellite DNA can undergo decondensation to a certain level. This process is accompanied by demethylation and transcription.

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Prolonged culture of normal chorionic villus cells yields ICF syndrome-like chromatin decondensation and rearrangements

Cited by 26 publications

References 38 publications

Specific hypermethylation of LINE-1 elements during abnormal overgrowth and differentiation of human placenta

Specific hypermethylation of LINE-1 elements during abnormal overgrowth and differentiation of human placenta

Chromosome Preparation From Cultured Cells

Human chromosome 1 satellite 3 DNA is decondensed, demethylated and transcribed in senescent cells and in A431 epithelial carcinoma cells

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