Phenotypic change and altered protein expression in X-ray and methylcholanthrene-transformed C3H10T1/2 fibroblasts.
The morphology, growth properties and cellular protein patterns from parent and two transformed C3H10T1/2 cell lines were analyzed to associate the phenotypic and protein differences with cell transformation. Transformed 10T1/2 cells were obtained by colony isolation after exposure of parent 10T1/2 cells to methylcholanthrene (MCA-1 cell line) or X-ray irradiation (XR-III cell line). Compared to parent 10T1/2 and MCA-1 cells, XR-III cells were much smaller in size and exhibited the highest growth rate, greatest cell saturation density, increased plating efficiency and greater expression of proliferating cell nuclear antigen. MCA-1 cells showed intermediate characteristics between parent and XR-III cells. Among the three cell lines, only XR-III cells showed anchorage-independent growth in soft agar. When [35S]methionine-labeled whole cell lysate proteins were separated by two-dimensional polyacrylamide gel electrophoresis, computer comparison algorithms revealed a 97% similarity in protein profiles among almost 800 proteins detected from each cell line. However, comparison of proteins patterns of the transformed cell lines to that of parent 10T1/2 cells showed that 30 and 20 proteins were induced or repressed in XR-III cells and MCA-1 cells, respectively. Similarly, 81 and 24 proteins showed significant quantitative changes (threefold or greater) in XR-III and MCA-1 cells, respectively, as compared with parent 10T1/2 cell proteins. The anchorage-independent growth and increased proliferation properties of XR-III cells suggest a later stage of transformation compared to MCA-1 cells.(ABSTRACT TRUNCATED AT 250 WORDS)
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Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709.
Merrick
B A
BA
Witcher
L L
LL
Patterson
R M
RM
Daluge
D R
DR
Selkirk
J K
JK
D000362:Agar; D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis; D002455:Cell Division / Q000502:physiology; D002461:Cell Line, Transformed / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D008748:Methylcholanthrene; D051379:Mice; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D010641:Phenotype; D011233:Precipitin Tests; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis; D014965:X-Rays
D000951:Antigens, Neoplasm; D009363:Neoplasm Proteins; D018809:Proliferating Cell Nuclear Antigen; D008748:Methylcholanthrene; D000362:Agar