This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Welch, D. R. Articles by Alessandrini, A. Search for Related Content PubMed PubMed Citation Articles by Welch, D. R. Articles by Alessandrini, A.

Transfection of Constitutively Active Mitogen-activated Protein/Extracellular Signal-regulated Kinase Kinase Confers Tumorigenic and Metastatic Potentials to NIH3T3 Cells¹

Jake Gittlen Cancer Research Institute, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033-2390 [D. R. W., T. S., T. O. L., S. F. G., D. J. H.]; Medical Services, Massachusetts General Hospital, Charlestown, Massachusetts [R. P., J. V. B., A. A.]; Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts [Q. H., R. L. E.]; and Instituto Venezolano Investigaciones Cientificas, Caracas, Venezuela [M. R., M. S. R.]

Cellular growth and differentiation are controlled by multiple extracellular signals, many of which activate extracellular signal-regulated kinase (ERK)/mitogen-activated protein (MAP) kinases. Components of the MAP kinase pathways also cause oncogenic transformation in their constitutively active forms. Moreover, expression of activated ras can confer metastatic potential upon some cells. Activation of MAP kinases requires phosphorylation of both Thr and Tyr in the catalytic domain by a family of dual-specificity kinases, called MEKs (MAP kinase/ERK kinase). MEK1 is activated by phosphorylation at Ser²¹⁸ and Ser²²² by Raf. Mutation of these two sites to acidic residues, specifically [Asp²¹⁸], [Asp²¹⁸, Asp²²²], and [Glu²¹⁸, Glu²²²], results in constitutively active MEK1. Using these mutant variants of MEK1, we showed previously that transfection of NIH/3T3 or Swiss 3T3 cells causes morphological transformation and increases growth on soft agar, independent of ERK activity. The transformed cell lines show increased expression of matrix metalloproteinases 2 and 9 and cathepsin L, proteinases that have been implicated in the metastatic process. We tested NIH3T3 cells transfected with the [Asp²¹⁸] or [Asp²¹⁸, Asp²²²] for metastatic potential after i.v. injection into athymic mice. Parental 3T3 cells formed no tumors grossly or histologically. However, all MEK1 mutant transformants formed macroscopic metastases. Thus, like activated Ras, MEK1 can confer both tumorigenic and metastatic potential upon NIH3T3 cells. These results refine the mechanism through which ras could confer tumorigenic and metastatic potential (i.e., the critical determinants of tumorigenic and metastatic potential are downstream of MEK1).

This article has been cited by other articles:

				D. E. Discher, P. Janmey, and Y.-l. Wang Tissue Cells Feel and Respond to the Stiffness of Their Substrate Science, November 18, 2005; 310(5751): 1139 - 1143. [Abstract] [Full Text] [PDF]

				N. Gavert, M. Conacci-Sorrell, D. Gast, A. Schneider, P. Altevogt, T. Brabletz, and A. Ben-Ze'ev L1, a novel target of {beta}-catenin signaling, transforms cells and is expressed at the invasive front of colon cancers J. Cell Biol., February 14, 2005; 168(4): 633 - 642. [Abstract] [Full Text] [PDF]

				J. Pinkas, S. S. Martin, and P. Leder Bcl-2-Mediated Cell Survival Promotes Metastasis of EpH4 {beta}MEKDD Mammary Epithelial Cells Mol. Cancer Res., October 1, 2004; 2(10): 551 - 556. [Abstract] [Full Text] [PDF]

				M. M. Schuierer, F. Bataille, S. Hagan, W. Kolch, and A.-K. Bosserhoff Reduction in Raf Kinase Inhibitor Protein Expression Is Associated with Increased Ras-Extracellular Signal-Regulated Kinase Signaling in Melanoma Cell Lines Cancer Res., August 1, 2004; 64(15): 5186 - 5192. [Abstract] [Full Text] [PDF]

				S. Silletti, M. Yebra, B. Perez, V. Cirulli, M. McMahon, and A. M. P. Montgomery Extracellular Signal-regulated Kinase (ERK)-dependent Gene Expression Contributes to L1 Cell Adhesion Molecule-dependent Motility and Invasion J. Biol. Chem., July 9, 2004; 279(28): 28880 - 28888. [Abstract] [Full Text] [PDF]

				B. Bedogni, M. S. O'Neill, S. M. Welford, D. M. Bouley, A. J. Giaccia, N. C. Denko, and M. B. Powell Topical Treatment with Inhibitors of the Phosphatidylinositol 3'-Kinase/Akt and Raf/Mitogen-Activated Protein Kinase Kinase/Extracellular Signal-Regulated Kinase Pathways Reduces Melanoma Development in Severe Combined Immunodeficient Mice Cancer Res., April 1, 2004; 64(7): 2552 - 2560. [Abstract] [Full Text] [PDF]

				T. Takino, H. Miyamori, Y. Watanabe, K. Yoshioka, M. Seiki, and H. Sato Membrane Type 1 Matrix Metalloproteinase Regulates Collagen-Dependent Mitogen-Activated Protein/Extracellular Signal-Related Kinase Activation and Cell Migration Cancer Res., February 1, 2004; 64(3): 1044 - 1049. [Abstract] [Full Text] [PDF]

				D. J. Vander Griend and C. W. Rinker-Schaeffer A New Look at an Old Problem: The Survival and Organ-Specific Growth of Metastases Sci. Signal., January 20, 2004; 2004(216): pe3 - pe3. [Abstract] [Full Text] [PDF]

				C.-C. Hung, T. Ichimura, J. L. Stevens, and J. V. Bonventre Protection of Renal Epithelial Cells against Oxidative Injury by Endoplasmic Reticulum Stress Preconditioning Is Mediated by ERK1/2 Activation J. Biol. Chem., August 1, 2003; 278(31): 29317 - 29326. [Abstract] [Full Text] [PDF]

				D. R. Welch and K. W. Hunter A New Member of the Growing Family of Metastasis Suppressors Identified in Prostate Cancer J Natl Cancer Inst, June 18, 2003; 95(12): 839 - 841. [Full Text] [PDF]

				Z. Fu, P. C. Smith, L. Zhang, M. A. Rubin, R. L. Dunn, Z. Yao, and E. T. Keller Effects of Raf Kinase Inhibitor Protein Expression on Suppression of Prostate Cancer Metastasis J Natl Cancer Inst, June 18, 2003; 95(12): 878 - 889. [Abstract] [Full Text] [PDF]

				J. Pinkas and P. Leder MEK1 Signaling Mediates Transformation and Metastasis of EpH4 Mammary Epithelial Cells Independent of an Epithelial to Mesenchymal Transition Cancer Res., August 15, 2002; 62(16): 4781 - 4790. [Abstract] [Full Text] [PDF]

				A. K. Greenberg, S. Basu, J. Hu, T.-a. Yie, K. M. Tchou-Wong, W. N. Rom, and T. C. Lee Selective p38 Activation in Human Non-Small Cell Lung Cancer Am. J. Respir. Cell Mol. Biol., May 1, 2002; 26(5): 558 - 564. [Abstract] [Full Text] [PDF]

				H. Mahloogi, D. E. Bassi, and A. J.P. Klein-Szanto Malignant conversion of non-tumorigenic murine skin keratinocytes overexpressing PACE4 Carcinogenesis, April 1, 2002; 23(4): 565 - 572. [Abstract] [Full Text] [PDF]

				D. Wang and A. Richmond Nuclear Factor-kappa B Activation by the CXC Chemokine Melanoma Growth-stimulatory Activity/Growth-regulated Protein Involves the MEKK1/p38 Mitogen-activated Protein Kinase Pathway J. Biol. Chem., January 26, 2001; 276(5): 3650 - 3659. [Abstract] [Full Text] [PDF]

				O. P. Barry, B. Mullan, D. Sheehan, M. G. Kazanietz, F. Shanahan, J. K. Collins, and G. C. O'Sullivan Constitutive ERK1/2 Activation in Esophagogastric Rib Bone Marrow Micrometastatic Cells Is MEK-independent J. Biol. Chem., April 27, 2001; 276(18): 15537 - 15546. [Abstract] [Full Text] [PDF]