[CANCER RESEARCH 53. 4376-4.1K2. September 15. 199.1]
Chemotaxis and Haptotaxis of Human Malignant Mesothelioma Cells:
Effects of Fibronectin, Laminin, Type IV Collagen, and an Autocrine
Motility Factor-like Substance1
Julius Klominek,2 Karl-Henrik Robert, and Karl-Costa Sundqvist
De/hirlment of Lung Medium1 /./. K.¡,Medicine [K-H. R.j. unii Clinical lininiinolog\\
[J. K,, K-(!. S.[. Karolinska Institute til Huililin^e University Hospital,
S-Ì41Kt>Huddinge. Sweden
ABSTRACT
A human malignant pleural mesothelioma cell line (STAY) was studied
with respect to production of the extracellular matrix components
lumi
nili, type IV collagen, and fibronectin, and interactions with these proteins
in vitro. We also analyzed S'l'AV cell serum-free
conditioned medium with
respect to the possible presence of "autocrine motility
factor-like" sub
stance. Sodium dodecylsulfale-polyacrylamide
gel electrophoresis
of bio-
synthetically
labeled STAY serum-free conditioned medium showed that
STAY cells released several proteins
into the medium,
including compo
nents with molecular weights of 850,000,
540,000 and 440.000. Using
Western blotting we identified these proteins as luminili, type IY collagen,
and fihronectin,
respectively. By immunocytochemistry
laminin,
type IY
collagen, and fibronectin were detected as a matrix surrounding
the cells.
Plastic culture dishes coated with /ug quantities of luminili, type IY colla
gen, and fibronectin
induced attachment
and spreading of STAY cells.
Laminin, type IY collagen, and fibronectin stimulated directional
(chemo-
tactic) migration of STAY cells
in Boyden chambers Pitted with 8 ¿im
filters. The same cells also migrated to insoluble step gradients of filter-
bound extracellular matrix components
(haptotaxis). When STAY serum-
free conditioned medium was separated by using fast protein liquid chro-
matography Superóse 6 gel filtration,
two motility-inducing
protein peaks
were detected. The first peak contained proteins with molecular weight >
220,000 that had both chemotactic
and haptot