Detection of genetic alterations in the human gastric cancer cell lines by two-dimensional analysis of genomic DNA.
We examined genomic DNA from each of three human-derived gastric cancer cell lines, using the technique of restriction landmark genomic scanning (RLGS) which allows monitoring of approximately 2, 000 NotI landmarks. The resulting DNA spots from cancer cell DNA were compared with those in normal mucosa or gastric primary tumor. In all, 9 intense spots were detected from two of the three cancer cell lines. Two highly intensified spots were common in the two cancer cell lines and proven to be originated from DNA region containing the human c-myc proto-oncogene on chromosome 8. The degree of amplification of c-myc DNA was similar to each other and was estimated to be 60-fold as compared to those from normal mucosa DNA. On the basis of chromosome-assigned RLGS (CA-RLGS), other spots were assigned to each chromosome, such as one on chromosome 8, two each from chromosome 20, and three on one of chromosome 9-12. The remaining spot seems to be due to demethylation of a repetitive element. Twenty-four spot that were lost due to either homozygous deletion or methylation on corresponding NotI cleavage sites were commonly observed in all cancer cells. These spots were also assigned to each chromosome: one each from chromosome 2, 6, 7, 13, 14, 16, and 20, two each from chromosome 3 and 5, and nine from chromosome 9-12 by CA-RLGS. Many of the multi-copy spots corresponding to ribosomal RNA genes were greatly decreased due mainly to methylation on CpG islands along with minor rDNA variants, indicating that only minor rRNA genes may be silenced in these cancer cells. These results show that the present alterations detected by RLGS might be useful for identification of candidate genes inactivated or expressed unexpectedly in tumor development and tumor progression in the stomach.
Kim
Y S
YS
Genome Research Center, KRIBB, Yusung, Taejon 305-600, Korea.
Yoo
H S
HS
Lee
K T
KT
Goh
S H
SH
Jung
J S
JS
Oh
S W
SW
Baba
M
M
Yasuda
T
T
Matsubara
K
K
Nagai
H
H
D001483:Base Sequence; D004273:DNA, Neoplasm / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics*; D005753:Gastric Mucosa / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D000090063:Proto-Oncogene Mas; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA / Q000379:methods*; D013274:Stomach Neoplasms / Q000235:genetics*; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D004275:DNA, Ribosomal; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas