Identification of CpG islands hypermethylated in human lung cancer by the arbitrarily primed-PCR method.
DNA hypermethylation is believed to be involved in human carcinogenesis, since it suppresses the transcription of defined genes and is associated with chromosomal instability. In this study, we identified CpG islands that are hypermethylated in human lung cancer by a modified arbitrarily primed-polymerase chain reaction method using genomic DNAs digested with a methylation-sensitive restriction enzyme, HpaII, as templates. When we analyzed genomic DNAs from normal lung tissues and non-small cell lung carcinoma cell lines using three arbitrary primers, three DNA fragments were amplified from lung cancer DNAs but not from normal lung DNAs. Restriction mapping and Southern blot analysis revealed that all of these bands were amplified from CpG islands that were hypermethylated in the lung cancer cell lines. These islands were mapped to chromosomes 4q34, 10q26 and 17p13.1-p13.2, respectively, and these chromosomal regions were also hypermethylated in a subset of primary lung tumors in vivo. Thus, diverse chromosomal regions are hypermethylated in lung cancer cells. The results also indicate that this method is simple and effective for screening of CpG islands that are hypermethylated in cancer cells.
Kohno
T
T
Biology Division, National Cancer Center Research Institute, Tokyo, Japan.
Kawanishi
M
M
Inazawa
J
J
Yokota
J
J
D002277:Carcinoma / Q000235:genetics*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D018899:CpG Islands / Q000235:genetics*; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000032:analysis; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm