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Originally published In Press as doi:10.1074/jbc.M001837200 on May 11, 2000

J. Biol. Chem., Vol. 275, Issue 30, 23326-23332, July 28, 2000
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Prolactin-induced Cell Proliferation in PC12 Cells Depends on JNK but Not ERK Activation*

Yu Cheng, Igor Zhizhin, Robert L. Perlman, and Dimitra MangouraDagger

From the Kennedy Center, Department of Pediatrics, Committee on Neurobiology and Committee Cell Physiology, University of Chicago, Chicago, Illinois 60637

The effects of pituitary and extrapituitary prolactin include cellular proliferation and differentiation. PC12 cells was used as a model to delineate respective signaling of prolactin. Prolactin acted as a mitogen for undifferentiated PC12 cells, as measured by significant increases in bromodeoxyuridine incorporation and in cell numbers, with an efficacy equal to epidermal growth factor. Both the long and short form of the prolactin receptor was expressed, yet only the long isoform was tyrosine-phosphorylated upon agonist binding. Functional prolactin receptor signaling was further demonstrated in the activation of JAK2 and phosphorylation activation of the transcription factors Stat1, -3, and -5a. Surprisingly, prolactin stimulated a sustained activation of Raf-B, without activation of the MAP kinases ERK1 or -2. Instead, in solid phase kinase assays using a glutathione S-transferase-c-Jun fusion protein (amino acids 1-79) as the substrate, a significant activation of the mitogen-activated protein Janus kinase (c-Jun N-terminal kinase; JNK) was observed. The prolactin-induced activation of JNK was prolonged and accompanied by a significant increase in c-Jun mRNA abundance and c-Jun protein synthesis. Moreover, analysis of bromodeoxyuridine incorporation at the single cell level revealed that epidermal growth factor-dependent incorporation was inhibited by PD98059 and independent of SB203580, whereas prolactin-induced incorporation was ERK and mitogen-activated protein kinase p38 independent but was abolished with JNK inhibition by 30 µM SB203580. Our studies suggest that prolactin may have a role in the growth of PC12 cells, where it stimulates concurrent mitogenic and differentiation-promoting signaling pathways.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Dept. of Pediatrics, Kennedy Center, MC 5058, WCH C-576, Chicago, IL 60637-1470. Tel.: 773-702-1136; Fax: 773-702-9234; E-mail: dm36@midway.uchicago.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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