Quantitative microarray-based DNA methylation analysis of E-cadherin gene promoter in acute leukemia.
- Author:
Bao-an CHEN
1
;
Fan ZHANG
;
Yan WANG
;
Wen-li ZHENG
;
Juan DU
;
Chong GAO
;
Jia-hua DING
;
Yun-yu SUN
;
Jian CHENG
;
Jun WANG
;
Gang ZHAO
;
Ning-na CHEN
;
Zu-hong LU
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Cadherins; genetics; CpG Islands; genetics; DNA Methylation; Humans; Leukemia, Myeloid, Acute; genetics; Molecular Sequence Data; Oligonucleotide Array Sequence Analysis; methods; Precursor Cell Lymphoblastic Leukemia-Lymphoma; genetics; Promoter Regions, Genetic
- From: Chinese Journal of Oncology 2007;29(1):41-44
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo quantitatively detect the methylation of E-cadherin gene 5'-CpG islands in acute leukemia by microarray-based DNA analysis and to briefly discuss the role of microarry for detection of methylation in tumors.
METHODSBisulfite-modified DNA was used as a template for PCR amplification, resulting in conversion of unmethylated cytosine, but not methylated cytosine, into thymine within CpG islands of interest. Five sets of oligonucleotide probes were designed to fabricate a DNA microarray to detect the methylation changes of E-cadherin gene CpG islands in acute leukemia. By drawing a standard curve to assess the levels of changes in methylation detected in the examined samples.
RESULTSMicroarray assay was successfully used to quantitatively detect methylation changes of E-cadherin gene in 5 acute leukemia samples. Varying degree of methylation was detected in five regions and the hypermethylation region was the same. The result was validated by gene sequencing.
CONCLUSIONMicroarray assay may be applied as an useful tool for mapping methylation changes in multiple CpG loci and for leukemia research. It is more time-saving and labor-saving than gene sequencing and can be used to quantitatively detect changes in methylation with high throughput.