[Quantitative microarray-based DNA methylation analysis of E-cadherin gene promoter in acute leukemia].
OBJECTIVE
To 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.


METHODS
Bisulfite-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.


RESULTS
Microarray 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.


CONCLUSIONS
Microarray 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.
Chen
Bao-an
BA
The Affiliated Zhongda Hospital of Southeast University, Nanjing 210009, China. bachen@seu.edu.cn
Zhang
Fan
F
Wang
Yan
Y
Zheng
Wen-li
WL
Du
Juan
J
Gao
Chong
C
Ding
Jia-hua
JH
Sun
Yun-yu
YY
Cheng
Jian
J
Wang
Jun
J
Zhao
Gang
G
Chen
Ning-na
NN
Lu
Zu-hong
ZH
D001483:Base Sequence; D015820:Cadherins / Q000235:genetics*; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics*; D011401:Promoter Regions, Genetic
D015820:Cadherins