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We investigated the mechanism of action of the human prolactin
(hPRL) receptor on four different breast cancer cell lines, T-47D,
MCF-7, BT-474, and SK-BR3, that express elevated levels of the receptor
compared with normal cells. Cells treated with human growth hormone
(hGH), which binds and activates the hPRL receptor, exhibited
bell-shaped dose-response growth curves consistent with the sequential
dimerization mechanism proposed for the hPRL receptor (Fuh, G., Colosi,
P., Wood, W.I., and Wells, J.A.(1993) J. Biol. Chem. 268,
5376-5381). Growth stimulation was enhanced by
Zn
Volume 270,
Number 22,
Issue of June 2, pp. 13133-13137, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
, which preferentially increases the affinity of
hGH for the hPRL receptor. Furthermore, receptor-selective variants of
hGH that bind the hPRL receptor but not the hGH receptor were
agonistic, providing additional support that specific binding to the
hPRL receptor can stimulate these breast cancer cells to grow. On this
basis we produced variants of hGH and human placental lactogen (hPL)
that were potential antagonists because they bind but do not dimerize
the hPRL receptor. The hPL-based antagonist was less potent than the
hGH-based antagonist toward the growth of MCF-7 cells, consistent with
the lower affinity of hPL for hPRL receptor than for hGH. However, the
hPL-based antagonist was more potent than the hGH antagonist for BT-474
cells. Antibodies to the hPRL receptor inhibited growth of FDC-P1 cells
transfected with the hPRL receptor; these also inhibited MCF-7 cells
and T47D cells but not BT-474 cells. A unique feature of BT-474 cells
was found when screening its cDNA revealed the presence of a novel
alternative splice of the hPRL receptor that codes for the soluble
extracellular domain; this may explain these differential inhibitory
effects. These studies provide further molecular insight into the
potential role of the hPRL receptor in breast cancer and demonstrate
that hPRL receptor antagonists can inhibit the growth of breast cancer
cells.
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