PCR detection of retinoblastoma gene deletions in radiation-induced mouse lung adenocarcinomas

Conference · Fri Jan 01 04:00:00 EST 1993

OSTI ID:6552291

Churchill, M E; Gemmell, M A; Woloschak, G E

From 1971 to 1986, Argonne National Laboratory conducted a series of large-scale studies of tumor incidence in 40,000 BCF[sub 1] mice irradiated with [sup 60]Co [gamma] rays or JANUS fission-spectrum neutrons; normal and tumor tissues from mice in these studies were preserved in paraffin blocks. A polymerase chain reaction (PCR) technique has been developed to detect deletions in the mouse retinoblastoma (mRb) gene in the paraffin-embedded tissues. Microtomed sections were used as the DNA source in PCR reaction mixtures. Six mRb gene exon fragments were amplified in a 40-cycle, 3-temperature PCR protocol. The absence of any of these fragments (relative to control PCR products) on a Southern blot indicated a deletion of that portion of the mRb gene. The tumors chosen for analysis were lung adenocarcinomas that were judged to be the cause of death in post-mortem analyses. Spontaneous tumors as well as those from irradiated mice (569 cGy of [sup 60]Co [gamma] rays or 60 cGy of JANUS neutrons, doses that have been found to have approximately equal biological effectiveness in the BCF, mouse) were analyzed for mRb deletions. In all normal mouse tissues studies, all six mRb exon fragments were present on Southem blots. Tumors in six neutron-irradiated mice also had no mRb deletions. However, I of 6 tumors from [gamma]-irradiated mice and 6 of 18 spontaneous tumors from unirradiated mice had a deletion in one or both mRb alleles. All deletions detected were in the 5[prime] region of the mRb gene.

• Research Organization: Argonne National Lab., IL (United States)
• Sponsoring Organization: DOE; USDOE, Washington, DC (United States)
• DOE Contract Number: W-31109-ENG-38
• OSTI ID: 6552291
• Report Number(s): ANL/BIM/CP-79174; CONF-930454--1; ON: DE93010659
• Country of Publication: United States
• Language: English

Related Subjects

550900 -- Pathology
560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
ANIMALS
BARYONS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARCINOGENESIS
CELL TRANSFORMATIONS
COBALT 60
COBALT ISOTOPES
DNA SEQUENCING
ELECTROMAGNETIC RADIATION
ELEMENTARY PARTICLES
FERMIONS
GAMMA RADIATION
GENE AMPLIFICATION
GENE MUTATIONS
GENES
HADRONS
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
IONIZING RADIATIONS
ISOMERIC TRANSITION ISOTOPES
ISOTOPES
LUNGS
MAMMALS
MICE
MINUTES LIVING RADIOISOTOPES
MUTANTS
MUTATIONS
NEUTRONS
NUCLEI
NUCLEONS
ODD-ODD NUCLEI
ONCOGENES
ONCOGENIC TRANSFORMATIONS
ORGANS
PATHOGENESIS
RADIATION INDUCED MUTANTS
RADIATIONS
RADIOISOTOPES
RESPIRATORY SYSTEM
RESPIRATORY TRACT CELLS
RODENTS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
YEARS LIVING RADIOISOT