Morphological alteration of X-ray induced partially transformed human cells by transfection with a small c-myc DNA sequence.
During attempts to transform a normal human fibroblast strain (GM730) by X-irradiation, we obtained a partially transformed cell strain (GM730pt) which demonstrates several aspects of the transformed phenotype including morphological changes, increased saturation density, growth in soft agar, and focus formation in long-term cultures. When GM730pt cells were transfected with the feline c-myc gene, morphology of the cells changed dramatically following seven days of expression. Transfection of other plasmid DNAs or oncogenes such as pUC8, pSV2neo, src, sis, and H-ras had little or no effects on the phenotype of GM730pt cells. On the other hand, a gel purified, small fragment of c-myc DNA had a complete cell alteration activity. Furthermore, Bal 31 deletion and M13 sequencing experiments showed that the alteration seen in GM730pt cells is delimited to a 24 nucleotide stretch (active myc element) from the second intron of the feline c-myc gene that contains a T-rich sequence.
Kano
Y
Y
Department of Health Science, Kibi International University, Okayama, Japan.
Nohno
T
T
Takahashi
R
R
Hiragami
F
F
Kawamura
K
K
Strebhardt
K
K
Namba
M
M
Sugiyama
T
T
Little
J B
JB
D000362:Agar; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002415:Cats; D002452:Cell Count / Q000528:radiation effects; D002455:Cell Division / Q000528:radiation effects; D002461:Cell Line, Transformed; D048429:Cell Size / Q000528:radiation effects; D002471:Cell Transformation, Neoplastic / Q000235:genetics* / Q000473:pathology / Q000528:radiation effects*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology* / Q000528:radiation effects*; D016259:Genes, myc / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D007231:Infant, Newborn; D007438:Introns / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009857:Oncogenes / Q000235:genetics / Q000502:physiology; D010957:Plasmids / Q000235:genetics / Q000502:physiology; D011071:Poly T / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics; D013997:Time Factors; D014162:Transfection; D014410:Tumor Stem Cell Assay
D011071:Poly T; D000362:Agar