Epigenetic reprogramming of the human <i>H19</i> gene in mouse embryonic cells does not erase the primary parental imprint

Mitsuya, Kohzoh; Meguro, Mitsue; Sui, Hajime; Schulz, Thomas C.; Kugoh, Hiroyuki; Hamada, Hiroshi; Oshimura, Mitsuo

doi:10.1046/j.1365-2443.1998.00183.x

Cited by 22 publications

(20 citation statements)

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“…We analysed the methylation status of the di erentially methylated HpaII sites (CCGG) within the KvDMR1 locus by Southern blot and by a HpaII-duplex-PCR. Southern blot hybridization with probe AA155639 (Mitsuya et al, 1999) of genomic DNA digested with the restriction enzymes BamHI and HpaII can distinguish the 6.0 kb fully methylated maternal allele from the 2.6 kb non-methylated paternal allele ( Figure 1a). Initially, we assessed the KvDMR1 methylation status of 11 normal somatic tissues derived from skeletal muscle, kidney, liver, stomach, lung, breast, ovary, bladder, uterus, placenta and peripheral blood.…”

Section: Analysis Of Kvdmr1 Methylation In Adult and Childhood Tumorsmentioning

confidence: 99%

“…Genomic DNAs from tumor samples were doubledigested with BamHI and HapII, a methylation-sensitive restriction enzyme. Hybridization was performed with probe AA155639 (Mitsuya et al, 1999) (a) The ®gure shows representative selected results from hybridizations of liver (b), cervical (d), breast (e), colorectal carcinomas (f), Wilms' tumors (c) and normal tissues (g). Control for lack of inhibition of restriction digestion was obtained by re-hybridization of blots shown in panels e and f with a STIM-1 probe (Sabbioni et al, 1999) (h,i).…”

Section: Loss Of Heterozygosity At 11p15mentioning

confidence: 99%

“…The KvDMR1 probe, corresponding to the EST AA155639 (Mitsuya et al, 1999), was ampli®ed by polymerase chain reaction (PCR) from placenta DNA using primers LIT1-A (5'-CCA AAA CCT CTC TGA AAT CTG-3') and LIT1-B (5'-GAT AAG GAG TAG GCC AAG GAT G-3'). The H19 probe 1, for the analysis of methylation of the H19 upstream region (Frevel et al, 1999) was ampli®ed from the BAC CITB-142F11 with primers H19C-9526F (5'-GAC TCT GTC CTG CGG AAA C-3') and H19D-9886R (5'-GAG ACA GGG CTG AGC ATT G-3').…”

Section: Dna Probesmentioning

confidence: 99%

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Loss of methylation at chromosome 11p15.5 is common in human adult tumors

Scelfo

Schwienbacher

Veronese³

et al. 2002

Oncogene

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Chromosome 11p15 deletion is frequent in human tumors, suggesting the presence of at least one tumor suppressor gene within this region. While mutation analyses of local genes revealed only rare mutations, we have previously described a mechanism, gain of imprinting, that leads to loss of expression of genes located on the maternal 11p15 chromosome in human hepatocarcinomas. Loss of expression was often associated with loss of maternal-speci®c methylation at the KvDMR1 locus. Here, we show that loss of the maternal KvDMR1 methylation is common, ranging from 30 to 50%, to a variety of adult neoplasms, including liver, breast, cervical and gastric carcinomas. We found that other 11p15.5 loci were concomitantly hypomethylated, indicating that loss of KvDMR1 methylation occurred in the context of a common mechanism a ecting the methylation of a large 11p15 subchromosomal domain. These epigenetic abnormalities were not detected in any normal somatic tissue. Therefore, it seems possible that, contrary to the repression of promoter activity caused by hypermethylation, loss of gene expression at 11p15.5 may result from the activation, by hypomethylation, of one or more negative regulatory elements.

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Section: Analysis Of Kvdmr1 Methylation In Adult and Childhood Tumorsmentioning

confidence: 99%

Section: Loss Of Heterozygosity At 11p15mentioning

confidence: 99%

Section: Dna Probesmentioning

confidence: 99%

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Loss of methylation at chromosome 11p15.5 is common in human adult tumors

Scelfo

Schwienbacher

Veronese³

et al. 2002

Oncogene

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“…We have constructed a library of mouse A9 hybrids containing single human paternal or maternal chromosomes via microcell-mediated chromosome transfer . The parent-of-originspecific expression and DNA methylation of human imprinted genes, such as H19, SNRPN, and IPW, were maintained faithfully (Meguro et al 1997;Mitsuya et al 1998) as was the replication timing of H19 (unpublished data) in these A9 hybrids.…”

Section: Introductionmentioning

confidence: 89%

“…The imprinted human genes, LIT1 and H19 on chromosome 11 and SNRPN on chromosome 15, were selected for analysis of allelic histone H4 acetylation using the ChIP assay (Crane-Robinson et al 1999). In the A9 hybrids, the epigenetic status of the transferred human chromosome is maintained faithfully; for example, the expression pattern is dependent on the parental origin and the DNA methylation status (Meguro et al 1997;Mitsuya et al 1998Mitsuya et al , 1999Kugoh et al 1999).…”

Section: Histone H4 Acetylation Of Imprinted Genesmentioning

confidence: 99%

A novel in vitro system for analyzing parental allele-specific histone acetylation in genomic imprinting

2001

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One of the obstacles in studying human genomic imprinting is distinguishing the parental origin of alleles in diploid cells. To solve this problem, we have constructed a library of mouse A9 hybrids in which individual clones contain a single human chromosome of known parental origin. Here we extend this in vitro system to the analysis of the role of histone acetylation in the allelic expression of human imprinted genes. The levels of histone H4 acetylation of the imprinted human LIT1, H19, and SNRPN genes were examined by a chromatin immunoprecipitation (ChIP) assay in mouse A9 hybrids with a single human chromosome of known parental origin. We demonstrated that H4 histones associated with the actively expressed alleles of imprinted LIT1, H19, and SNRPN genes were highly acetylated, whereas they were hypoacetylated in the silent alleles. Furthermore, treatment of A9 hybrids with trichostatin A (TSA), an inhibitor of histone deacetylase, resulted in transcriptional reactivation of the silent alleles for LIT1 and SNRPN, suggesting that histone deacetylation is one of the key regulatory mechanisms in genomic imprinting. These results indicate that our monochromosomal hybrid system is a new technology for analyzing histone modifications between parental alleles in human imprinted genes.

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Epigenetic silencing of PEG3 gene expression in human glioma cell lines*

Maegawa

Yoshioka

Itaba

et al. 2001

Molecular Carcinogenesis

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Genomic imprinting, the phenomenon in which alleles of genes are expressed differentially depending on their parental origins, has important consequences for mammalian development, and disturbance of normal imprinting leads to abnormal embryogenesis and some inherited diseases and is also associated with various cancers. In the context of screening for novel imprinted genes on human chromosome 19q13.4 with mouse A9 hybrids, we identified a maternal allele-specific methylated CpG island in exon 1 of paternally expressed imprinted gene 3 (PEG3), a gene that exhibits paternal allele-specific expression. Because PEG3 expression is downregulated in some gliomas and glioma cell lines, despite high-level expression in normal brain tissues, we investigated whether the loss of PEG3 expression is related to epigenetic modifications involving DNA methylation. We found monoallelic expression of PEG3 in all normal brain tissues examined and five of nine glioma cell lines that had both unmethylated and methylated alleles; the remaining four glioma cell lines exhibited gain of imprinting with hypermethylated alleles. In addition, treatment of glioma cell lines with the DNA demethylating agent 5-aza-2'-deoxycytidine reversed the silencing of PEG3 biallelically. In this article, we report that the epigenetic silencing of PEG3 expression in glioma cell lines depends on aberrant DNA methylation of an exonic CpG island, suggesting that PEG3 contributes to glioma carcinogenesis in certain cases.

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Epigenetic reprogramming of the human H19 gene in mouse embryonic cells does not erase the primary parental imprint

Cited by 22 publications

References 50 publications

Loss of methylation at chromosome 11p15.5 is common in human adult tumors

Loss of methylation at chromosome 11p15.5 is common in human adult tumors

A novel in vitro system for analyzing parental allele-specific histone acetylation in genomic imprinting

Epigenetic silencing of PEG3 gene expression in human glioma cell lines*

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