Characterization of the plasminogen activator of herpesvirus-transformed cells and examination of its correlation with the tumorigenic and metastatic ability of in vivo-derived sublines
Herpes simplex virus type 2-transformed hamster embryo fibroblasts (333-8-9 cells) produce increased amounts of plasminogen activator (PA) compared with normal hamster cells. The 333-8-9 PA activity was quantitated in comparison to a PA standard, urokinase (UK). Using a direct PA assay in which /sup 125/I-labeled plasminogen is cleaved, a linear dose-response was seen over a 1000-fold range in UK concentration when plotted on a semi-logarithmic scale. Extracellular PA activity secreted by the HSV-2-transformed cell line, 333-8-9, followed a similar dose-response slop. The optimum pH and osmolarity for detection of the 333-8-9 extracellular PA activity were pH 8.9 and approximately 150 mOsmol, respectively. Secretion of PA by the 333-8-9 cells did not vary significantly on a per cell basis over cell densities ranging from 0.1 to 8.0 x 10/sup 7/ cells/T-75 cm/sup 2/ flask. This assay was accurate, reproducible, and demonstrated that the 333-8-9 cells produced at least a 20-fold greater amount of PA activity than their normal cell counterparts. Based on the molecular weight (50-58 Kd) of the secreted 333-8-9 cell PA and lack of fibrin stimulation of the PA activity, it is concluded to be a urokinase-type PA.
- Research Organization:
- Pennsylvania State Univ., University Park (USA)
- OSTI ID:
- 5243657
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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UROKINASE
BIOCHEMICAL REACTION KINETICS
ENZYME ACTIVITY
DOSE-RESPONSE RELATIONSHIPS
EMBRYOS
FIBROBLASTS
HERPES SIMPLEX
IODINE 125
MOLECULAR WEIGHT
PLASMINOGEN
TRACER TECHNIQUES
ANIMAL CELLS
BETA DECAY RADIOISOTOPES
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DISEASES
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
FIBRINOLYTIC AGENTS
HEMATOLOGIC AGENTS
HYDROLASES
INFECTIOUS DISEASES
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
NONSPECIFIC PEPTIDASES
NUCLEI
ODD-EVEN NUCLEI
PEPTIDE HYDROLASES
RADIOISOTOPES
REACTION KINETICS
SKIN DISEASES
SOMATIC CELLS
VIRAL DISEASES
550201* - Biochemistry- Tracer Techniques