[Application of microarrays in screening the tumor-specific genes in the genome of K562 cells].
OBJECTIVE
To screen tumor-specific genes of K562 cells using DNA microarray technique.


METHODS
The genomic DNA of normal white blood cells and cultured K562 cells were respectively purified and digested with Sau3A I, and the digested DNA fragments of K562 cells were cloned into TA cloning vector to construct the corresponding genomic DNA library. The insert genomic DNA fragments were amplified from the library to prepare the microarray using Cartesian 5500 Microarrayer. The digested genomic DNA fragments of normal white blood cells were labeled with fluorescent Cy3 by restriction display PCR (RD-PCR), followed by hybridization with the microarray, after which the slide was washed and scanned with ScanArray.


RESULTS
Among the 426 target genes, 42 differential genes were identified in the genomic DNA of K562 cells in comparison with the normal white blood cells. One of the genes was identified as the breakpoint cluster gene (BCR) after sequence analysis.


CONCLUSIONS
The DNA microarrays we constructed may effectively identify the tumor-specific genes in K562 cells, and DNA microarray technique can be helpful in elucidating the molecular mechanisms of tumorigenesis at the genomic level.
Peng
Gui-fu
GF
Institute of Molecular Biology, First Military Medical University, Guangzhou 510515, China.
Ma
Wen-li
WL
Song
Yan-bing
YB
Peng
Yi-fei
YF
Shi
Rong
R
Mao
Xiang-ming
XM
Zheng
Wen-ling
WL
D000595:Amino Acid Sequence; D016678:Genome; D006801:Humans; D020014:K562 Cells; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics*; D007962:Leukocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*