Neoplastic transformation of mouse C3H10T1/2 cells following exposure to neutrons does not involve mutation of ras gene as analyzed by SSCP and cycle sequencing.
About 25% of human tumors contain a mutated member of the ras gene family. Neutron exposure is an occupational risk in several work places and while we know that cells exposed to neutrons can become transformed, the molecular basis of this process is not understood. To determine whether neutron-induced cellular transformation involves ras mutation, C3H10T1/2 cells were exposed to a single dose of 5.9 MeV neutrons. Type II and type III foci were isolated and established as cell lines. A total of 34 foci were selected and expanded for analysis of tumorigenicity, chromosomal aberrations and mutations in members of the ras gene family. The presence of mutations in genomic DNA in N-ras or K-ras of each focus was examined by either single-strand conformational polymorphism (SSCP) analysis or by asymmetric PCR coupled cell cycle sequence analysis. Although chromosomal aberrations were detected at metaphase, no alterations in either ras gene were detected. We conclude that in vitro neutron-induced transformation must occur through a mechanism other than ras mutation.
Freyer
G A
GA
Center for Radiological Research, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
Palmer
D A
DA
Yu
Y
Y
Miller
R C
RC
Pandita
T K
TK
D000818:Animals; D001483:Base Sequence; D002470:Cell Survival / Q000528:radiation effects; D002471:Cell Transformation, Neoplastic / Q000528:radiation effects*; D002869:Chromosome Aberrations; D011905:Genes, ras; D051379:Mice; D008809:Mice, Inbred C3H; D009502:Neutrons; D018807:Polymorphism, Single-Stranded Conformational