PCR detection of retinoblastoma gene deletions in radiation-induced mouse lung adenocarcinomas
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:
- USDOE, Washington, DC (United States)
- DOE Contract Number:
- W-31109-ENG-38
- OSTI ID:
- 10145607
- Report Number(s):
- ANL/BIM/CP-79174; CONF-930454-1; ON: DE93010659
- Resource Relation:
- Conference: International seminar on molecular mechanisms in radiation mutagenesis and carcinogenesis,Doorwerth (Netherlands),19-22 Apr 1993; Other Information: PBD: [1993]
- Country of Publication:
- United States
- Language:
- English
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PCR detection of retinoblastoma gene deletions in radiation-induced mouse lung adenocarcinomas
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59 BASIC BIOLOGICAL SCIENCES
GENE MUTATIONS
DNA SEQUENCING
RESPIRATORY TRACT CELLS
ONCOGENIC TRANSFORMATIONS
ONCOGENES
GENE AMPLIFICATION
RADIATION INDUCED MUTANTS
CARCINOGENESIS
LUNGS
MICE
COBALT 60
GAMMA RADIATION
NEUTRONS
560120
550900
RADIATION EFFECTS ON BIOCHEMICALS, CELLS, AND TISSUE CULTURE
PATHOLOGY