Expression of O6-methylguanine-DNA methyltransferase in six human medulloblastoma cell lines.
Six well characterized human medulloblastoma cell lines (D283 Med, Daoy, D341 Med, D384 Med, D425 Med, and D458 Med) were examined for the expression of O6-methylguanine-DNA methyltransferase (MGMT) by activity and Western and Northern blot analysis. High levels of MGMT activity were present in D283 Med, Daoy, D341 Med, and D384 Med (1.36, 0.80, 1.68, and 1.62 pmol/mg of protein, respectively), but negligible MGMT activity was detected in D425 Med and D458 Med (0.06 and 0.05 pmol/mg of protein, respectively), which were derived separately at different times from the same patient. The presence of MGMT protein and its transcript was demonstrated in D283 Med, Daoy, D341 Med, and D384 Med, but both the protein and the mRNA were undetectable in D425 Med and D458 Med. Nevertheless, all six cell lines contained an apparently unaltered MGMT gene, as determined by Southern blot analysis. The absence of MGMT activity in D425 Med and D458 Med is likely due to the absence of the protein, resulting from a lack of transcription of the MGMT gene. The varying levels of expression of MGMT in medulloblastoma cells found in this study should provide a molecular basis for drug design and selection in chemotherapy of this tumor.
He
X M
XM
Department of Pathology, Duke University Medical Center, Durham, North Carolina 27710.
Ostrowski
L E
LE
von Wronski
M A
MA
Friedman
H S
HS
Wikstrand
C J
CJ
Bigner
S H
SH
Rasheed
A
A
Batra
S K
SK
Mitra
S
S
Brent
T P
TP
D015152:Blotting, Northern; D015153:Blotting, Western; D006801:Humans; D008527:Medulloblastoma / Q000201:enzymology*; D008780:Methyltransferases / Q000378:metabolism*; D019853:O(6)-Methylguanine-DNA Methyltransferase; D012333:RNA, Messenger / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D008780:Methyltransferases; D019853:O(6)-Methylguanine-DNA Methyltransferase