Receptors for d-Trp 6 -Luteinizing Hormone-Releasing Hormone,Somatostatin,and Insulin-Like Growth Factor I in MXT Mouse Mammary Carcinoma

Abstract

Abstract

Membrane receptors for d-Trp⁶-luteinizing hormone-releasing hormone (d-Trp⁶-LH-RH), somatostatin-14 (SS-14), and insulin-like growth factor I (IGF-I) were estimated in MXT mammary cancers of mice using sensitive multipoint micromethods. The receptors were characterized in untreated animals and following in vivo treatment with microcapsules of the agonist d-Trp⁶-LH-RH and the somatostatin analog RC-160, which strongly inhibited tumor growth. In the control group, d-Trp⁶-LH-RH was bound to the single class of saturable, specific, noncooperative receptor sites (K_d , = 29.3 ± 8.48 × 10^-9 M; B_max = 4.55 ± 0.31 pmol/mg membrane protein). Treatment with d-Trp⁶-LH-RH alone or in combination with RC-160 produced down-regulation of membrane receptors for d-Trp⁶-LH-RH on MXT mammary tumor cells. RC-160 alone and ovariectomy were without effect on d-Trp⁶-LH-RH receptors. On the membrane surface of MXT mammary cells, we found one class of high affinity, specific, saturable binding sites for SS-14 (K_d = 4.4 ± 1.9 × 10^-9 M; B_max = 0.58 ± 0.21 pmol/mg membrane protein). Treatment with RC-160 alone or combined with d-Trp⁶-LH-RH significantly increased both the dissociation binding constant (K _d = 18.6 ± 3.5 × 10^-9 and 10.1 ± 0.7 × 10^-9 M, respectively) and the binding capacity (B_max = 13.98 ± 1.7 and 21.00 ± 4.0 pmol/mg membrane protein, respectively). We also found specific binding sites (K _d = 3.01 ± 0.15 × 10^-9 M; B_max = 2.24 ± 0.96 pmol/mg membrane protein) for IGF-I in the membrane fractions of MXT mammary cancers. Chronic treatment with d-Trp⁶-LH-RH and RC-160 alone or in combination, as well as ovariectomy, significantly decreased the dissociation binding constant of IGF-I membrane receptors on MXT mammary cells. Our results strongly suggest an important role of LH-RH, SS-14, and IGF-I in the growth of MXT mammary carcinoma. Changes in characteristics of receptors after treatment with analogs of LH-RH and SS-14 along with tumor growth inhibition provide additional support for the direct effect of these peptides on tumor cells. A possible significance of these findings as applied to a clinical environment is discussed.