A study on the relationship between antisense EGFR cDNA fragments and nuclear matrix proteins in glioblastoma cells.
The association of antisense epidermal growth factor receptor (EGFR) cDNA fragments with nuclear matrix from EGFR-antisense transfected glioblastoma cell lines U343 and U87 was investigated. A 1015 bp DNA fragment (primer I-II) was amplified in both genomic DNA and nuclear matrix-associated DNA (NM DNA) from EGFR-antisense transfected glioblastoma cell lines U343E and U87E. Two different DNA fragments (940 bp and 110 bp) were amplified by primer I-III in both genomic DNA and NM DNA of U343E, while one 110 bp PCR product was shown with the same primer in both genomic DNA and NM DNA of U87E only. After EGFR-antisense transfection, the binding property of the 110 bp DNA fragment (primer IV-V) to nuclear matrix was not affected. Southwestern blotting demonstrated the presence of antisense EGFR cDNA binding nuclear matrix proteins. Our findings demonstrate that not only EGFR DNA is associated with nuclear matrix, but the transfected antisense EGFR cDNA also binds to nuclear matrix proteins. The nuclear matrix is most likely involved in the replication and transcription of antisense EGFR cDNA or hybridisation with sense mRNA in vitro.
Wang
Z H
ZH
Department of Anatomy, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, PR China.
Yam
G H
GH
Tian
X X
XX
Ng
H K
HK
Chew-Cheng
S B
SB
Ding
M X
MX
Chew
E C
EC
D001932:Brain Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000378:metabolism*; D066246:ErbB Receptors / Q000235:genetics*; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D034521:Nuclear Matrix-Associated Proteins / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D014162:Transfection
D018076:DNA, Complementary; D034521:Nuclear Matrix-Associated Proteins; D016376:Oligonucleotides, Antisense; C512478:EGFR protein, human; D066246:ErbB Receptors