Genomic Screen for Epigenetically Silenced Genes Associated With Cancer
a gene and gene technology, applied in the field of gene detection methods, can solve the problems of patients not seeking medical assistance, and the lack of markers for most cancers, and achieve the effect of convenient detection and facilitation of detection
Inactive Publication Date: 2009-04-16
THE JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE
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- Description
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- Application Information
AI Technical Summary
Benefits of technology
"The present invention relates to methods for identifying epigenetically silenced genes associated with cancer, particularly methylation silenced genes. These methods involve contacting a genome with nucleic acid subtraction products that selectively hybridize to RNA molecules expressed in cancer cells but not in normal cells. The nucleic acid subtraction products can be nucleic acid molecules corresponding to RNA expressed in cancer cells or nucleic acid molecules that reactivate expression of epigenetically silenced genes. The methods can be performed using methyltransferase inhibitors, histone deacetylase inhibitors, or a combination of both. The epigenetically silenced genes can be associated with various types of cancer, such as carcinoma or sarcoma."
Problems solved by technology
Although cancers generally are considered to be due to genetic changes such as mutations of a gene, it has become clear that epigenetic mechanisms, which do not result in mutations of the DNA sequence, also can result in cancers.
Unfortunately, such markers are not available for most cancers.
As such, many cancer patients do not seek medical assistance until the cancer is at a stage that requires radical therapy, or is untreatable.
Method used
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Examples
Experimental program
Comparison scheme
Effect test
example 1
Genomic Screen for Epigenetic Silenced Gene Associated with Colorectal Cancer
[0120]This example provides a method for detecting genes that are epigenetically down-regulated in cancer cells, and confirms the validity of the method by identifying genes that are epigenetically down-regulated in and diagnostic of colorectal cancer cells (see, also, Suzuki et al., Nature Genet. 31:141-149, 2002, which is incorporated herein by reference).
Methods
Cell Culture and Tissue Samples
[0121]Cell lines were cultured in RPMI 1640 or Minimal Essential Medium (MEM) supplemented with 10% fetal bovine serum. Tissue samples of colorectal cancer and normal colon mucosa were from specimens obtained at the time of clinically indicated surgical procedures.
DAC and TSA Treatment and RNA Preparation
[0122]RKO cells were treated with 5-aza-2′-deoxycytidine (DAC; Sigma) and / or trichostatin A (TSA; Wako) as described (Cameron et al., supra, 1999). Briefly, the treatment consisted of DAC (200 mM) for 48 hr, with dru...
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Abstract
A method of identifying epigenetically silenced genes, e.g., methylation silenced genes, in cancer cells is provided. In addition, methods of identifying a cancer by detecting epigenetic silencing of gene expression are provided, as are methods of treating a subject having such a cancer, for example, a colorectal cancer and / or gastric cancer. Reagents for practicing such methods also are provided.
Description
[0001]This application is a continuation of U.S. Ser. No. 10 / 384,491, filed Mar. 7, 2003 (pending), which claims the benefit of priority under 35 U.S.C. § 119(e)(1) of U.S. Ser. No. 60 / 362,422, filed Mar. 7, 2002, the entire content of which is incorporated herein by reference.[0002]This invention was made in part with government support under Grant No. CA54396 awarded by the National Cancer Institute. The United States government has certain rights in this invention.BACKGROUND OF THE INVENTION[0003]1. Field of the Invention[0004]The invention relates generally to methods of detecting genes that are epigenetically silenced in cancer cells, and more specifically to a genomic screen useful for identifying colorectal cancer cells and gastric cancer cells.[0005]2. Background Information[0006]Although cancers generally are considered to be due to genetic changes such as mutations of a gene, it has become clear that epigenetic mechanisms, which do not result in mutations of the DNA sequen...
Claims
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IPC IPC(8): C12Q1/68C40B30/06A61K31/7068G01N33/53A61K38/46A61K45/00A61K48/00A61P35/00C12M1/00C12N15/09C12Q1/02C12Q1/34G01N37/00
CPCC12Q1/6809C12Q1/6886C12Q2600/154C12Q2537/164A61P35/00
Inventor BAYLIN, STEPHEN B.HERMAN, JAMESSUZUKI, HIROMU
Owner THE JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE