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J Biol Chem, Vol. 274, Issue 26, 18428-18437, June 25, 1999
152/135) BOUND WITH
Sp1
From the Department of Cancer Biology, M.D. Anderson Cancer Center,
Houston, Texas 77030
Since c-src overexpression increases
colonic cell invasiveness and because both Src activity and urokinase
receptor protein are elevated in invasive colon cancers, the present
study was undertaken: 1) to determine if a constitutively active Src
regulates urokinase receptor expression and 2) to identify required
cis-elements and trans-acting factors. SW480
colon cancer cells transfected with an expression plasmid (c-srcY527F)
encoding a constitutively active Src protein manifested increased
urokinase receptor gene expression and Src activity. Treatment of the
src transfectants with a Src-inhibitor (PD173955) reduced
urokinase receptor protein levels and laminin degradation. Inasmuch as
we recently implicated an upstream region of the urokinase receptor
promoter (152/135) in constitutive urokinase receptor expression,
we determined its role for the induction by src. Whereas
the activity of a CAT reporter driven by this region was stimulated by
c-srcY527F, the u-PAR promoter mutated at the Sp1-binding motif in the
152/135 region was not. Nuclear extracts from the src
transfectants demonstrated increased Sp1 binding to region 152/135
compared with those from SW480 cells. Finally, endogenous urokinase
receptor protein amounts in 10 colon cancers and corresponding normal
colon correlated with Src specific activity. These data suggest that
urokinase receptor gene expression is regulated by Src partly via
increased Sp1 binding.
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