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Geriatric Research, Education, and Clinical Center, VA Puget Sound Health Care System (S.E.D., L.M., S.R.P.), Tacoma, Washington 98493; Division of Gerontology and Geriatric Medicine, Department of Medicine, University of Washington (S.E.D., L.M., S.R.P.), Seattle, Washington; and the Department of Pathology (J.L.W.), Medical College of Virginia, Richmond, Virginia 23298
Address all correspondence and requests for reprints to: Stephen Plymate, Geriatric Research, Education, and Clinical Center, VA Puget Sound Health Care System, American Lake Division, Tacoma, Washington 98493. E-mail: splymate{at}u.washington.edu
Insulin-like growth factor (IGF) binding proteins (IGFBPs) have been
shown to either inhibit or enhance the action of IGF, or act in an
IGF-independent manner in the prostate. We have overexpressed the
IGF-inhibitory IGFBP-4 in the malignant M12 prostate epithelial cell
line to determine the effects on tumor formation and apoptosis.
Overexpression was determined by Northern, Western immunoblot and
Western radioligand blot analysis. IGF-induced proliferation was
reduced in the IGFBP-4 transfected cells compared with control cells
(P 
0.01). Colony formation in soft agar was
significantly inhibited up to 14 days after plating in the IGFBP-4
transfected cells when compared with the M12 controls
(P 0.01): however, in the presence of
des(1–3)IGF-I, there was no significant difference between the control
and IGFBP-4 transfectants in colony formation in soft agar. Apoptosis
in an IGFBP-4 transfected cell line was significantly increased in
response to induction by 6-hydroxyurea compared with the control line.
When injected sc into male athymic/nude mice, a marked delay was noted
in tumor formation in animals receiving IGFBP-4 transfected cells
(P 0.01). Interestingly, IGFBP-2 protein levels
were reduced in the conditioned media of all IGFBP-4 transfected cell
cultures. These data indicate that an inhibitory IGFBP may
significantly delay the growth of malignant prostate epithelial cells
and enhance the sensitivity of these cells to apoptosis.
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