Up-regulation of lysyl oxidase in spontaneous revertants of H-ras-transformed rat fibroblasts.
Neoplastic transformation mediated by ras oncogenes is associated with down-regulation of gene expression. We have constructed a subtracted complementary DNA library from preneoplastic rat 208F fibroblasts by hybridizing with mRNA from a ras-transformed subclone. One of the complementary DNA clones identified by this approach encodes the 3' end of lysyl oxidase, the homologue of the mouse ras recision gene. Expression of lysyl oxidase was almost completely down-regulated in two clones of H-ras-transformed 208F cells (FE-8 and FE-56). We isolated a set of spontaneous phenotypic revertants of FE-8 cells (designated FSR) by cloning at limiting dilution. FSR revertant clones expressed high levels of lysyl oxidase and H-ras mRNA but grew only poorly in semisolid agar medium as opposed to anchorage-independent parental FE-8 cells. We obtained subclones of FSR cells which displayed again the transformed morphology of FE-8 cells but required anchorage for growth and continued to express high levels of lysyl oxidase mRNA. Thus, expression of lysyl oxidase correlated with the suppression of anchorage-independent growth rather than with flat morphology. Lysyl oxidase might be a useful marker to distinguish between different aspects of reversion and transformation.
Hajnal
A
A
Department of Pathology, University Hospital, University of Zurich, Switzerland.
Klemenz
R
R
Schäfer
R
R
D000465:Algorithms; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002461:Cell Line, Transformed / Q000201:enzymology; D002471:Cell Transformation, Neoplastic; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D011905:Genes, ras; D008969:Molecular Sequence Data; D011061:Poly A / Q000378:metabolism; D016133:Polymerase Chain Reaction; D008249:Protein-Lysine 6-Oxidase / Q000096:biosynthesis*; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000378:metabolism; D051381:Rats; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D008249:Protein-Lysine 6-Oxidase