Not hMSH2 but hMLH1 is frequently silenced by hypermethylation in endometrial cancer but rarely silenced in pancreatic cancer with microsatellite instability.

Kondo, Emiko; Furukawa, Toru; Yoshinaga, Kousuke; Kijima, Hiroshi; Semba, Shuho; Yatsuoka, Toshimasa; Yokoyama, Toshiharu; Fukushige, Shinichi; Horii, Akira

doi:10.3892/ijo.17.3.535

Cited by 35 publications

(30 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Antibodies used for chromatin immunoprecipitation were anti-normal rabbit immunoglobulin G (Calbiochem), anti-MBD4 (H300), anti-MBD2 (Upstate), antidimethyl-histone H3 (Lys9) clone RR202 (Upstate), anti-acetyl-histone H3 (Upstate), and anti-acetyl-histone H4 (Upstate). PCR amplification was performed in 15 l with the p16 INK4a CpG island primers (18) and with the hMLH1 primers (14).…”

Section: Methodsmentioning

confidence: 99%

The Thymine DNA Glycosylase MBD4 Represses Transcription and Is Associated with Methylated p16 ^INK4a and hMLH1 Genes

et al. 2005

View full text Add to dashboard Cite

Epigenetic silencing through methyl-CpG (mCpG) is implicated in many biological patterns such as genome imprinting, X chromosome inactivation, and cancer development. In this process, the mCpG binding domain (MBD) proteins play an essential role in transmitting epigenetic information to downstream regulatory proteins. Among the five MBD proteins identified so far, MBD4 has been the only exception; it has long been thought to be a DNA repair protein. Herein we demonstrate that MBD4 has the ability to repress transcription through mCpG. Transcriptional repression by the MBD4 is histone deacetylase (HDAC) dependent, and MBD4 directly binds to Sin3A and HDAC1 at three central regions that overlap transcriptional repression domains. Furthermore, a chromatin immunoprecipitation assay clearly shows that MBD4 binds to hypermethylated promoters of the p16INK4a and hMLH1 genes. These results suggest that MBD4 is one of the essential components involved in epigenetic silencing in cancer and its repair activity is necessary for the maintenance of hypermethylated promoters.

show abstract

Section: Methodsmentioning

confidence: 99%

The Thymine DNA Glycosylase MBD4 Represses Transcription and Is Associated with Methylated p16 ^INK4a and hMLH1 Genes

et al. 2005

View full text Add to dashboard Cite

show abstract

“…8 The antigen retrieval method employed for staining of all molecules but DUSP6 were as described. 28 The antibodies employed were a polyclonal anti-DUSP6 antibody, C-20 (Santa Cruz Biotechnology Lab Inc., Santa Cruz, CA, USA), monoclonal anti-CDKN2A antibodies, G175-405 (BD Biosciences, San Jose, CA, USA) and DCS-50 (SIGMA, St Louis, MO, USA), a monoclonal anti-TP53 antibody, DO-7 (Dako Cytomation Co. Ltd, Glostrup, Denmark), a monoclonal anti-SMAD4 antibody, B8 (Santa Cruz Biotechnology Lab Inc.), a monoclonal anti-MUC1-CORE antibody, Ma552 (Novocastra Laboratories Ltd. Newcastle upon Tyne, UK), a monoclonal anti-MUC2 antibody, Ccp58 (Santa Cruz Biotechnology Lab Inc.), and a monoclonal anti-MUC5AC antibody, CLH2 (Chemicon International Inc., Temecula, CA, USA) for primary antibody reactions and biotin-conjugated anti-goat or mouse IgG (H þ L) antibodies (Vector Lab. Inc., Burlingham, CA, USA) for secondary antibody reactions.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Distinct progression pathways involving the dysfunction of DUSP6/MKP-3 in pancreatic intraepithelial neoplasia and intraductal papillary-mucinous neoplasms of the pancreas

et al. 2005

View full text Add to dashboard Cite

DUSP6/MKP-3 is identified as a candidate tumor suppressor gene for pancreatic cancer. The aim of this study was to elucidate the roles of DUSP6 in the pancreatic carcinogenesis through the pancreatic intraepithelial neoplasia and/or intraductal papillary-mucinous neoplasms, both of which are considered to be precursor lesions of invasive carcinoma of the pancreas, by comparing with involvements of other major tumor suppressive pathways. Expressions of DUSP6, CDKN2A, TP53, and SMAD4 were investigated by immunohistochemistry in a total of 206 lesions of dysplastic ductal precursors and carcinomas retrieved from 52 pancreata with invasive ductal carcinomas and 51 of those with intraductal papillary-mucinous neoplasms. The intensity of staining was evaluated in lesions at different atypical grades and statistically compared among them. Mutations of KRAS2 were analyzed by methods of the allele-specific oligonucleotide hybridization and nucleotide sequencing. In pancreata with invasive ductal carcinomas, expressions of DUSP6 were abrogated exclusively in the invasive carcinoma cells in contrast to its fairly preserved expressions in pancreatic intraepithelial neoplasia. In pancreata with intraductal papillary-mucinous neoplasms, abrogated expressions of DUSP6 were observed in a relatively small fraction of intraductal adenoma/borderlines and intraductal carcinomas. Most of the intraductal adenoma/borderline lesions with abrogation of DUSP6 harbored mutations of KRAS2. None of the molecules was associated with each other in any grade of lesions. Morphological variations of papillae of the intraductal papillary-mucinous neoplasms were evaluated and analyzed for their associations with abrogations of the molecules, which resulted in finding of no significant associations. Our results suggest that the abrogation of DUSP6 is associated exclusively with progression from pancreatic intraepithelial neoplasia to the invasive ductal carcinoma while it is potentially associated with initiation of intraductal papillary-mucinous neoplasms with mutated KRAS2, which is independent of other major tumor suppressive pathways in both types of neoplasms.

show abstract

“…25,26 Expression of the hMSH2 gene was monitored as a positive control as the gene was demonstrated to be expressed in all parental cells. 27 Samples of 29 primary human pancreatic cancers as well as the corresponding normal pancreatic tissues were also used. These tissues were obtained at Tohoku University Hospital with informed consent and were reviewed by a board-approved pathologist.…”

Section: Rt-pcrmentioning

confidence: 99%

Transcriptional silencing of ETS-1 efficiently suppresses angiogenesis of pancreatic cancer

Lefter

Dima²,

Sunamura

et al. 2008

Cancer Gene Ther

Self Cite

View full text Add to dashboard Cite

In this study, we addressed the hypothesis that transcriptional suppression of erythroblastosis virus E26 oncogene homolog 1 (ETS-1) is an efficient therapeutic approach to pancreatic adenocarcinoma by investigating the effect of ETS-1 suppression in human pancreatic cancer cells. We accomplished this by using an adenoviral vector encoding only the DNA-binding domain of wild-type ETS-1 (ETS-1 dominant negative, ETS-1-DN). ETS-1-DN decreases ETS-1-binding by competing for its binding to DNA. Adenoviralmediated transfer of ETS-1-DN (adenoviral ETS-1-DN construct, AdETS-1-DN) into pancreatic tumor cell lines did not affect their proliferation rate in vitro but did significantly inhibit their in vivo growth in nude mice. Furthermore, to test the efficacy of ETS-1-DN in vivo, we injected the AdETS-1-DN into established human pancreatic adenocarcinomas grown in nude mice. This treatment significantly reduced tumor size as compared to saline injection, without any detectable side effects. Microvessel density in mouse xenografts displayed significantly lower values in tumors in which ETS-1 was downregulated. In addition, expression of the ETS-1-DN in the pancreatic cancer cells resulted in downregulation of urokinase-type plasminogen activator (u-PA) and metalloproteinase-1 (MMP-1) expression. Taken together, these data suggest that transcriptional inactivation of ETS-1 is able to significantly affect angiogenesis and growth of pancreatic cancer. This effect may be due in part to downregulation of MMP-1 and u-PA expression. Our results suggest that ETS-1-DN is a promising candidate for antiangiogenic gene therapy in pancreatic cancer.

show abstract

Not hMSH2 but hMLH1 is frequently silenced by hypermethylation in endometrial cancer but rarely silenced in pancreatic cancer with microsatellite instability.

Cited by 35 publications

References 0 publications

The Thymine DNA Glycosylase MBD4 Represses Transcription and Is Associated with Methylated p16 ^INK4a and hMLH1 Genes

The Thymine DNA Glycosylase MBD4 Represses Transcription and Is Associated with Methylated p16 ^INK4a and hMLH1 Genes

Distinct progression pathways involving the dysfunction of DUSP6/MKP-3 in pancreatic intraepithelial neoplasia and intraductal papillary-mucinous neoplasms of the pancreas

Transcriptional silencing of ETS-1 efficiently suppresses angiogenesis of pancreatic cancer

Contact Info

Product

Resources

About

Not hMSH2 but hMLH1 is frequently silenced by hypermethylation in endometrial cancer but rarely silenced in pancreatic cancer with microsatellite instability.

Cited by 35 publications

References 0 publications

The Thymine DNA Glycosylase MBD4 Represses Transcription and Is Associated with Methylated p16 INK4a and hMLH1 Genes

The Thymine DNA Glycosylase MBD4 Represses Transcription and Is Associated with Methylated p16 INK4a and hMLH1 Genes

Distinct progression pathways involving the dysfunction of DUSP6/MKP-3 in pancreatic intraepithelial neoplasia and intraductal papillary-mucinous neoplasms of the pancreas

Transcriptional silencing of ETS-1 efficiently suppresses angiogenesis of pancreatic cancer

Contact Info

Product

Resources

About

The Thymine DNA Glycosylase MBD4 Represses Transcription and Is Associated with Methylated p16 ^INK4a and hMLH1 Genes

The Thymine DNA Glycosylase MBD4 Represses Transcription and Is Associated with Methylated p16 ^INK4a and hMLH1 Genes