Inhibition of DNA methyltransferase stimulates the expression of signal transducer and activator of transcription 1, 2, and 3 genes in colon tumor cells

Karpf, Adam R.; Peterson, Peter; Rawlins, Joseph T.; Dalley, Brian; Yang, Qian; Albertsen, Hans; Jones, David A.

doi:10.1073/pnas.96.24.14007

Cited by 174 publications

(136 citation statements)

References 44 publications

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“…One of the central themes we have synthesized from our microarray studies is that gene expression signatures are most easily understood as a representation of the activation status of cellular signal transduction pathways. An early example of this theme was our recognition that treatment of colon cancer cells with 5-aza-CdR leads to the activation of the interferon signal transduction pathway (Karpf et al, 1999). Our data support the notion that the induction of interferon pathway target genes following 5-aza-CdR treatment is an indirect effect, i.e.…”

Section: Mechanisms Leading To Gene Reactivationsupporting

confidence: 79%

Reactivating the expression of methylation silenced genes in human cancer

2002

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Section: Mechanisms Leading To Gene Reactivationsupporting

confidence: 79%

Reactivating the expression of methylation silenced genes in human cancer

2002

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“…Signal transducer and activator of transcription (STAT) -1, -2, and -3 genes were shown to be reactivated in tumor cells treated by 5-AzaC [6]. The STAT-3 gene has also been known to play some roles in maintaining undifferentiated status of primitive hematopoietic cells [7].…”

Section: Resultsmentioning

confidence: 99%

5‐azacytidine supports the long‐term repopulating activity of cord blood CD34⁺ cells

et al. 2004

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DNA methylation plays important roles in a wide range of biological phenomena, especially in the embryonic development and tumorigenesis. However, correlations between differentiation and DNA methylation have not been clarified well in each differentiation system. In this study, we focused our attention on regulatory roles of DNA methylation in normal hematopoietic differentiation using a demethylating reagent, 5-azacytidine (5-AzaC). As a source of hematopoietic progenitor cells, we used CD34 + cells prepared from human umbilical cord blood and examined the effects of 5-AzaC on the colony-forming activity and the long-term culture-initiating (LTC-IC) activity of these cells. 5-AzaC treatment increased LTC-IC frequency 1.57-to 2.50-fold as compared to the nontreated control. In parallel to this, immunoblotting analysis showed that the intensity of overall DNA methylation decreased after 5-AzaC treatment. These results indicated the involvement of DNA methylation and demethylation in controlling immaturity of hematopoietic progenitor cells and the usefulness of 5-AzaC for regulating this immaturity. Am.

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“…mRNA purified from RNAlater-treated tissues provided excellent templates for microarray analysis. B, Northern blot (performed as described previously [5]); lane 1, 1 g squamous cell carcinoma poly Aϩ-enriched RNA; lane 2, 1 g basal cell carcinoma poly Aϩ-enriched RNA (same RNA samples used on the microarray); the Northern blot was probed with a cDNA encoding the monokine induced by gamma interferon (MIG) that is also shown as a green spot on the microarray above. GAPDH expression is shown below as a loading control in panels B and C. Northern blot analysis confirms differential expression of MIG in these samples.…”

Section: Discussionmentioning

confidence: 99%

“…Oligotex (Qiagen Inc., Valencia, CA) was used to obtain poly Aϩ enriched RNA for use on the microarray. Poly Aϩ RNA was reverse transcribed and the BCC-derived RNA was labeled with Cy3-dCTP (green) while the SCCderived RNA was labeled with Cy5-dCTP (red) as previously published (5). The labeled cDNA was hybridized to a glass microscope slide harboring 4608 clones (in duplicate) and imaged by a Generation III Molecular Dynamics laser scanner (Sunnyvale, CA).…”

Section: Rna Preparation and Usementioning

confidence: 99%

Preservation of RNA for Functional Genomic Studies: A Multidisciplinary Tumor Bank Protocol

Coffin²,

et al. 2001

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Few human tumors are collected such that RNA is preserved for molecular analysis. Completion of the Human Genome Project will soon result in the identification of more than 100,000 new genes. Consequently, increasing attention is being diverted to identifying the function of these newly described genes. Here we describe a multidisciplinary tumor bank procurement protocol that preserves both the integrity of tissue for pathologic diagnosis, and the RNA for molecular analyses. Freshly excised normal skin was obtained from five patients undergoing wound reconstruction following Mohs micrographic surgery for cutaneous neoplasia. Tissues treated for 24 hours with RNAlater™ were compared histologically and immunohistochemically to tissues not treated with RNAlater. Immunohistochemical stains studied included: CD45, CEA, cytokeratin AE1/3, vimentin, S-100, and CD34 on formalin-fixed, paraffin embedded tissue and CD45 staining of frozen tissue. Slides were blinded and evaluated independently by three pathologists. The histologic and immunohistochemical parameters of tissue stored in RNAlater were indistinguishable from tissue processed in standard fashion with the exception of S-100 stain which failed to identify melanocytes or Langerhan's cells within the epidermis in any of the RNAlater -treated tissues. Interestingly, nerve trunks within the dermis stained appropriately for S-100. Multiple non-cutaneous autopsy tissues were treated with RNAlater, formalin, liquid nitrogen (LN2), and TRIzol Reagent . The pathologists were unable to distinguish between tissues treated with RNAlater, formalin, or frozen in LN2, but could easily distinguish tissues treated with TRIzol Reagent because of extensive cytolysis. RNA was isolated from a portion of the tissue treated with RNAlater and used for molecular studies including Northern blotting and microarray analysis. RNA was adequate for Northern blot analysis and mRNA purified from RNAlater -treated tissues consistently provided excellent templates for reverse transcription and subsequent microarray analysis. We conclude that tissues treated with RNAlater before routine processing are indistinguishable histologically and immunohistochemically from tissues processed in routine fashion and that the RNA isolated from these tissues is of high quality and can be used for molecular studies. Based on this study, we developed a multidisciplinary tumor bank procurement protocol in which fresh tissue from resection specimens are routinely stored in RNAlater at the time of preliminary dissection. Thus, precious human tissue can be utilized for functional genomic studies without compromising the tissue's diagnostic and prognostic qualities.

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Inhibition of DNA methyltransferase stimulates the expression of signal transducer and activator of transcription 1, 2, and 3 genes in colon tumor cells

Cited by 174 publications

References 44 publications

Reactivating the expression of methylation silenced genes in human cancer

Reactivating the expression of methylation silenced genes in human cancer

5‐azacytidine supports the long‐term repopulating activity of cord blood CD34⁺ cells

Preservation of RNA for Functional Genomic Studies: A Multidisciplinary Tumor Bank Protocol

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Inhibition of DNA methyltransferase stimulates the expression of signal transducer and activator of transcription 1, 2, and 3 genes in colon tumor cells

Cited by 174 publications

References 44 publications

Reactivating the expression of methylation silenced genes in human cancer

Reactivating the expression of methylation silenced genes in human cancer

5‐azacytidine supports the long‐term repopulating activity of cord blood CD34+ cells

Preservation of RNA for Functional Genomic Studies: A Multidisciplinary Tumor Bank Protocol

Contact Info

Product

Resources

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5‐azacytidine supports the long‐term repopulating activity of cord blood CD34⁺ cells