Invasive human fibrosarcoma DNA mediated induction of a 92 kDa gelatinase/type IV collagenase leads to an invasive phenotype.
Proteolytic enzymes, such as gelatinase/type IV collagenase, play a pivotal role in cancer invasion and metastasis. Invasive human fibrosarcoma cells (HT1080) secrete two species of gelatinase/type IV collagenase, 68-72 kDa and 92 kDa enzymes. The purpose of this study is to elucidate which species of gelatinase/type IV collagenase plays a more important role in invasion. We have found that HT1080 x human fibroblast hybrids have reduced ability to invade a reconstituted basement membrane (Matrigel) in vitro compared to HT1080 cells, and abundantly secrete only the 68-72 kDa gelatinase/type IV collagenase. These data suggest that the 92 kDa gelatinase/type IV collagenase may be more important in HT1080 cell invasion. We next transfected HT1080 genomic DNA into non-invasive mouse C3H/10T1/2 fibroblast cells, which secrete only 68-72 kDa gelatinase/type IV collagenase. Four invasive transfectants were established. These invasive transfectants secreted the 92 kDa gelatinase/type IV collagenase in addition to the 68-72 kDa gelatinase/type IV collagenase, whereas non-invasive control DNA transfectants did not secrete the 92 kDa gelatinase/type IV collagenase. These results suggest that the induction of the 92 kDa gelatinase/type IV collagenase is important in the invasive phenotype.
Kubota
S
S
Laboratory of Developmental Biology, National Institute of Dental Research, National Institutes of Health, Bethesda, MD 20892.
Mitsudomi
T
T
Yamada
Y
Y
D000818:Animals; D001483:Base Sequence; D001485:Basement Membrane / Q000502:physiology; D015139:Blotting, Southern; D002460:Cell Line; D004273:DNA, Neoplasm / Q000235:genetics* / Q000302:isolation & purification; D004790:Enzyme Induction; D005354:Fibrosarcoma / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D018093:Gelatinases; D011905:Genes, ras; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D008809:Mice, Inbred C3H; D003012:Microbial Collagenase / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009361:Neoplasm Invasiveness; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D010434:Pepsin A / Q000235:genetics*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D014162:Transfection
D004273:DNA, Neoplasm; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D010434:Pepsin A; D018093:Gelatinases; D003012:Microbial Collagenase