BIGEL analysis of gene expression in HL60 cells exposed to X rays or 60 Hz magnetic fields.
We screened a panel of 1,920 randomly selected cDNAs to discover genes that are differentially expressed in HL60 cells exposed to 60 Hz magnetic fields (2 mT) or X rays (5 Gy) compared to unexposed cells. Identification of these clones was accomplished using our two-gel cDNA library screening method (BIGEL). Eighteen cDNAs differentially expressed in X-irradiated compared to control HL60 cells were recovered from a panel of 1,920 clones. Differential expression in experimental compared to control cells was confirmed independently by Northern blotting of paired total RNA samples hybridized to each of the 18 clone-specific cDNA probes. DNA sequencing revealed that 15 of the 18 cDNA clones produced matches with the database for genes related to cell growth, protein synthesis, energy metabolism, oxidative stress or apoptosis (including MYC, neuroleukin, copper zinc-dependent superoxide dismutase, TC4 RAS-like protein, peptide elongation factor 1alpha, BNIP3, GATA3, NF45, cytochrome c oxidase II and triosephosphate isomerase mRNAs). In contrast, BIGEL analysis of the same 1,920 cDNAs revealed no differences greater than 1.5-fold in expression levels in magnetic-field compared to sham-exposed cells. Magnetic-field-exposed and control samples were analyzed further for the presence of mRNA encoding X-ray-responsive genes by hybridization of the 18 specific cDNA probes to RNA from exposed and control HL60 cells. Our results suggest that differential gene expression is induced in approximately 1% of a random pool of cDNAs by ionizing radiation but not by 60 Hz magnetic fields under the present experimental conditions.
Balcer-Kubiczek
E K
EK
Department of Radiation Oncology, School of Medicine; Veterans Affairs Hospital, Baltimore, Maryland 21201, USA.
Zhang
X F
XF
Han
L H
LH
Harrison
G H
GH
Davis
C C
CC
Zhou
X J
XJ
Ioffe
V
V
McCready
W A
WA
Abraham
J M
JM
Meltzer
S J
SJ
D004249:DNA Damage; D018076:DNA, Complementary / Q000235:genetics / Q000528:radiation effects; D015870:Gene Expression / Q000528:radiation effects*; D015723:Gene Library; D018922:HL-60 Cells; D006801:Humans; D008280:Magnetics / Q000009:adverse effects*; D009154:Mutation; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics
D018076:DNA, Complementary; D012333:RNA, Messenger