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Dual effect of prolactin on protein kinase C activity in CHO cells expressing functional prolactin receptors

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Journal of Biomedical Science

Abstract

We investigated the effects of prolactin (PRL) on the protein kinase C (PKC) activity in Chinese hamster ovary (CHO-E32) cells stably transfected with rabbit mammary gland PRL receptor cDNA. These cells express a functional long form of PRL-R. A 10-min to 2-hour treatment with 5 nM PRL resulted in the translocation of PKC activity from the cytosol to the membrane. Longer treatment (10–24 h) with the same concentration of PRL decreased the PKC activity in both particulate and cytoplasmic fractions. The PRL effect was dose dependent: maximal action was obtained with 1–10 nM. The PRL-induced activation of PKC was blocked by 20 nM staurosporine, a PKC inhibitor. Two inhibitors of tyrosine kinase, herbimycin A (1.75 µM) and genistein (100 µM), had no effect on PRL-induced activation of PKC.

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References

  1. Ali S, Pellegrini I, Kelly PA. A prolactin-dependent immune cell line (Nb2) expresses a mutant form of prolactin receptor. J Biol Chem 266:20110–20117;1991.

    PubMed  Google Scholar 

  2. Barros F, Katz GM, Kaczorowski GJ, Vandlen RL, Reuben JP. Calcium currents in GH3 cultured pituitary cells under whole-cell voltage-clamp: Inhibition by voltage-dependent potassium current. Proc Natl Acad Sci USA 82:1108–1112;1985.

    PubMed  Google Scholar 

  3. Boutin JM, Edery M, Shirota M, Jolicoeur C, Lesueur L, Ali S, Gould D, Djiane J, Kelly PA. Identification of a cDNA encoding a long form of prolactin receptor in human hepatoma and breast cancer cells. Mol Endocrinol 3:1455–1461;1989.

    PubMed  Google Scholar 

  4. Boutin J, Jolicoeur C, Okamura H, Cagnon J, Edery M, Shirota M, Banville D, Fourt I, Djiane J, Kelly PA. Cloning and expression of the rat prolactin receptor: A member of the growth hormone/prolactin receptor gene family. Cell 53:69–77;1988.

    Article  PubMed  Google Scholar 

  5. Bradford MM. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principal of protein-dye binding. Anal Biochem 72:248–254;1976.

    PubMed  Google Scholar 

  6. Buckley AR, Buckley DJ. Prolactin-stimulated ornithine decarboxylase induction in rat hepatocytes: Coupling to diacylglycerol generation and protein kinase C. Life Sci 48:237–243;1991.

    Article  PubMed  Google Scholar 

  7. Buckley AR, Crowe PD, Russell DH. Rapid activation of protein kinase C in isolated rat liver nuclei by prolactin, a known hepatic mitogen. Proc Natl Acad Sci USA 85:8649–8653;1988.

    PubMed  Google Scholar 

  8. Buckley AR, Montgomery DW, Kibler R, Putman CW, Zukoski C, Gout PW, Bee CT, Russell DH. Prolactin stimulation of ornithine decarboxylase and mitogenesis in Nb2 node lymphoma cells: The role of protein kinase C and calcium mobilization. Immunopharmacology 12:37–51;1986.

    Article  PubMed  Google Scholar 

  9. Doerner D, Abdel-Latif M, Rogers TB, Alger BE. Protein kinase C-dependent and -independent effects of phorbol esters on hippocampal calcium channel current. J Neurosci 10:1699–1706;1990.

    PubMed  Google Scholar 

  10. Doerner D, Pitler TA, Alger BE. Protein C activators block specific calcium and potassium current components in isolated hippocampal neurons. J Neurosci 8:4069–4078;1988.

    PubMed  Google Scholar 

  11. Dufy B, Jaken S, Barker JL. Ca2+-dependent PKC activation mimicks delayed effects of TRH on clonal pituitary cell excitability. Endocrinology 121:793–802;1987.

    PubMed  Google Scholar 

  12. Edery M, Jolicoeur C, Levi MC, Dusanter-Fourt I, Petridou B, Boutin J, Lesueur L, Kelly P, Djiane J. Identification and sequence analysis of a second form of prolactin receptor by molecular cloning of complementary DNA from rabbit mammary gland. Proc Natl Acad Sci USA 86:2112–2116;1989.

    PubMed  Google Scholar 

  13. Fan G, Rillema JA. Effect of a tyrosine kinase inhibitor, genistein, on the actions of prolactin in cultured mouse mammary tissues. Mol Cell Endocrinol 89:51–59;1992.

    Article  Google Scholar 

  14. Fukazawa H, Li PM, Yamamoto C, Murakami Y, Mizuno S, Uehara Y. Specific inhibition of cytoplasmic protein kinases by herbimycin A in vitro. Biochem Pharmacol 42:1661–1671;1991.

    Article  PubMed  Google Scholar 

  15. Gout PW, Beer CT, Noble RL. Prolactin-stimulated growth of cell cultures established from malignant Nb2 rat lymphoma. Cancer Res 40:2433–2436;1980.

    PubMed  Google Scholar 

  16. Hirota K, Hirota T, Aguilera G, Catt KJ. Hormone-induced redistribution of calcium-activated phospholipid-dependent protein kinase in pituitary gonadotrophs. J Biol Chem 260:3243–3246;1985.

    PubMed  Google Scholar 

  17. Kelly PA, Djiane J, Katoh M, Ferland LH, Houdebine LM, Teyssot B, Dusanter-Fourt I. The interaction of prolactin with its receptors in target tissues and its mechanism of action. Recent Prog Horm Res 40:379–438;1984.

    PubMed  Google Scholar 

  18. Kelly PA, Djiane J, Postel-Vinay MC, Edery M. The prolactin/growth hormone receptor family. Endocr/Rev 12:235–251;1991.

    Google Scholar 

  19. Ko KWS, Cook HW, Vance DE. Reduction of phosphatidylcholine turnover in Nb2 lymphoma cell line after prolactin treatment. J Biol Chem 261:7846–7852;1986.

    PubMed  Google Scholar 

  20. Kornberg LJ, Liberti JP. Lactogen-induced protein phosphorylation in Nb2 cells. Arch Biochem Biophys 279:109–115;1990.

    Article  PubMed  Google Scholar 

  21. Larsen JL, Dufau ML. Modulation of prolactin-stimulated Nb2 lymphoma cell mitogenesis by cholera toxin and pertussis toxin. Endocrinology 123:438–444;1988.

    PubMed  Google Scholar 

  22. Lebrun JJ, Ali S, Sofer L, Ullrich A, Kelly PA. Prolactin-induced proliferation of Nb2 cells involves tyrosine phosphorylation of the prolactin receptor and its associated tyrosine kinase JAK2. J Biol Chem 269:14021–14026;1994.

    PubMed  Google Scholar 

  23. Lesueur L, Edery M, Ali S, Paly J, Kelly PA, Djiane J. Comparison of long and short forms of the prolactin receptor on prolactin-induced milk protein gene transcription. Proc Natl Acad Sci USA 88:824–828;1991.

    PubMed  Google Scholar 

  24. Lesueur L, Edery M, Paly J, Clark J, Kelly PA, Djiane J. Prolactin stimulates milk protein promoter in CHO cells cotransfected with prolactin receptor cDNA. Mol Cell Endocrinol 71:7–12;1990.

    Article  Google Scholar 

  25. Nicoll CS, Bern HA. On the action of prolactin among the vertebrates: Is there a common denominator? In: Wolstenholme GEW, Knight J, eds. Lactogenic Hormones. London, Churchill-Livingstone, 299–327;1972.

    Google Scholar 

  26. Nishizuka Y. Studies and perspectives of protein kinase C. Science 233:305–312;1986.

    PubMed  Google Scholar 

  27. Parant MR, Vial HJ. Rapid and serial determination of protein kinase C activity and of the associated [3H]PDBu binding using a 96-well microtiter plate and a cell harvester. Anal Biochem 184:283–290;1990.

    Article  PubMed  Google Scholar 

  28. Prevarskaya N, Skryma R, Vacher P, Bignon C, Djiane J, Dufy B. Early effects of PRL on ion conductances in CHO cells expressing PRL receptor. Am J Physiol 267:C554-C562;1994.

    PubMed  Google Scholar 

  29. Rillema JA, Waters SB, Tarrant TM. Studies on the possible role of protein kinase C in the prolactin regulation of cell replication in Nb2 node lymphoma cells (42968). Proc Soc Exp Biol Med 192:140–144;1989.

    PubMed  Google Scholar 

  30. Rui H, Djeu JY, Evans GA, Kelly PA, Farrar W. Prolactin receptor triggering: Evidence for rapid tyrosine kinase activation. J Biol Chem 267:24076–24081;1992.

    PubMed  Google Scholar 

  31. Russell DH, Zorn NE, Buckley AR, Crowe PD, Sauro MD, Hadden EM, Farese RV, Laird HE. Prolactin and known modulators of rat splenocytes activate nuclear protein kinase C. Eur J Pharmacol 188:139–152;1990.

    Article  PubMed  Google Scholar 

  32. Sauro MD, Buckley AR, Russell DH, Fitzpatrick DF. Prolactin stimulation of protein kinase C activity in rat aortic smooth muscle. Life Sci 44:1787–1792;1989.

    Article  PubMed  Google Scholar 

  33. Shirota M, Banville D, Ali S, Jolicoeur C, Boutin M, Edery M, Djiane J, Kelly PA. Expression of two forms of prolactin receptor in rat ovary and liver. Mol Endocrinol 4:1136–1143;1990.

    PubMed  Google Scholar 

  34. Taupignon AI, Dufy B. Le phorbol myristate acétate favorise l'entrée de calcium dans les cellules hypophysaires GH3/B6 par les canaux calciques potentiel-dépendants. C R Soc Biol 182:74–78;1988.

    Google Scholar 

  35. Too CKL, Cragoe EJ Jr, Friesen HG. Amiloride-sensitive Na+/H+ exchange in rat Nb2 node lymphoma cells: Stimulation by prolactin and other mitogens. Endocrinolgy 121:1512–1520;1987.

    Google Scholar 

  36. Too CKL, Murphy PR, Friesen HG. G-proteins modulate prolactin-and interleukin-2-stimulated mitogenesis in rat Nb2 lymphoma cells. Endocrinology 124:2185–2192;1989.

    PubMed  Google Scholar 

  37. Too CKL, Shiu RPC, Friesen HG. Cross-linking of G-proteins to the prolactin receptor in rat Nb2 lymphoma cells. Biochem Biophys Res Commun 173:48–52;1990.

    Article  PubMed  Google Scholar 

  38. Too CKL, Walker A, Murphy P, Cragoe EJ Jr, Jacobs HK Jr, Friesen HG. Identification of amiloride-sensitive Na+/H+ exchange in rat Nb2 node lymphoma cells: Stimulation by 12-o-tetradecanoyl-phorbol-13-acetate. Endocrinology 121:1503–1511;1987.

    PubMed  Google Scholar 

  39. Uehara Y, Fukazawa H, Murakami Y, Mizuno S. Irreversible inhibition of V-src tyrosine kinase activity by herbimycin A and its abrogation by sulfhydryl compounds. Biochem Biophys Res Commun 163:803–809;1989.

    Article  PubMed  Google Scholar 

  40. Vacher P, Tran van Chuoi M, Paly J, Djiane J, Dufy B. Short term effect of prolactin on intracellular calcium in Chinese hamster ovary cells stably transfected with prolactin receptor complementary deoxyribonucleic acid. Endocrinology 134:1213–1218;1994.

    Article  PubMed  Google Scholar 

  41. Villalba M, Zabala MT, Martinez-Serrano A, De La Colina R, Satrustegui J, Garcia-Ruiz J. Prolactin increases cytosolic free calcium concentration in hepatocytes of lactating rats. Endocrinology 129:2857–2861;1991.

    PubMed  Google Scholar 

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Audy, M.C., Vacher, P., Vacher, A.M. et al. Dual effect of prolactin on protein kinase C activity in CHO cells expressing functional prolactin receptors. J Biomed Sci 3, 126–132 (1996). https://doi.org/10.1007/BF02255540

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