Exposure of murine cells to pulsed electromagnetic fields rapidly activates the mTOR signaling pathway

Murine pre‐osteoblasts and fibroblast cell lines were used to determine the effect of pulsed electromagnetic field (PEMF) exposure on the production of autocrine growth factors and the activation of early signal transduction pathways. Exposure of pre‐osteoblast cells to PEMF minimally increased the amount of secreted TGF‐β after 1 day, but had no significant effects thereafter. PEMF exposure of pre‐osteoblast cells also had no effect on the amount of prostaglandin E2 in the conditioned medium. Exposure of both pre‐osteoblasts and fibroblasts to PEMF rapidly activated the mTOR signaling pathway, as evidenced by increased phosphorylation of mTOR, p70 S6 kinase, and the ribosomal protein S6. Inhibition of PI3‐kinase activity with the chemical inhibitor LY294002 blocked PEMF‐dependent activation of mTOR in both the pre‐osteoblast and fibroblast cell lines. These findings suggest that PEMF exposure might function in a manner analogous to soluble growth factors by activating a unique set of signaling pathways, inclusive of the PI‐3 kinase/mTOR pathway. Bioelectromagnetics 27:535–544, 2006. © 2006 Wiley‐Liss, Inc.

[1]  T. Kubo,et al.  Electromagnetic Fields , 2008 .

[2]  Yoshitada Sakai,et al.  Pulsed electromagnetic field treatments enhance the healing of fibular osteotomies , 2005, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[3]  J. Blenis,et al.  Target of rapamycin (TOR): an integrator of nutrient and growth factor signals and coordinator of cell growth and cell cycle progression , 2004, Oncogene.

[4]  K. Inoki,et al.  TSC2 Mediates Cellular Energy Response to Control Cell Growth and Survival , 2003, Cell.

[5]  John C. Lee,et al.  Inhibition of phosphatidylinositol 3‐kinase and p70S6 kinase blocks osteogenic protein‐1 induction of alkaline phosphatase activity in fetal rat calvaria cells , 2003, Journal of cellular biochemistry.

[6]  L. Bonewald,et al.  Pulsed electromagnetic fields affect phenotype and connexin 43 protein expression in MLO‐Y4 osteocyte‐like cells and ROS 17/2.8 osteoblast‐like cells , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[7]  Maciej Zborowski,et al.  Magnetic Field Visualization in Applications to Pulsed Electromagnetic Field Stimulation of Tissues , 2003, Annals of Biomedical Engineering.

[8]  N. Kimura,et al.  A possible linkage between AMP‐activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) signalling pathway , 2003, Genes to cells : devoted to molecular & cellular mechanisms.

[9]  S. Pelech,et al.  Nocodazole-induced p53-dependent c-Jun N-terminal Kinase Activation Reduces Apoptosis in Human Colon Carcinoma HCT116 Cells* , 2002, The Journal of Biological Chemistry.

[10]  Dimitris J. Panagopoulos,et al.  Mechanism for action of electromagnetic fields on cells. , 2002, Biochemical and biophysical research communications.

[11]  Isao Ohnishi,et al.  Effect of pulsed electromagnetic fields (PEMF) on late‐phase osteotomy gap healing in a canine tibial model , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[12]  Neus Agell,et al.  Modulation of the Ras/Raf/MEK/ERK pathway by Ca(2+), and calmodulin. , 2002, Cellular signalling.

[13]  P. Lazarovici,et al.  Signaling Pathways for PC12 Cell Differentiation: Making the Right Connections , 2002, Science.

[14]  Roy K. Aaron,et al.  Upregulation of basal TGFβ1 levels by EMF coincident with chondrogenesis – implications for skeletal repair and tissue engineering , 2002 .

[15]  B. Caterson,et al.  Low frequency EMF regulates chondrocyte differentiation and expression of matrix proteins , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[16]  A. Jaeschke,et al.  Mammalian TOR: A Homeostatic ATP Sensor , 2001, Science.

[17]  O. Kozawa,et al.  Involvement of p70 S6 kinase in bone morphogenetic protein signaling: Vascular endothelial growth factor synthesis by bone morphogenetic protein‐4 in osteoblasts , 2001, Journal of cellular biochemistry.

[18]  A. Gingras,et al.  Regulation of translation initiation by FRAP/mTOR. , 2001, Genes & development.

[19]  J. Heckman,et al.  Pulsed Electromagnetic Fields Increase Growth Factor Release by Nonunion Cells , 2001, Clinical orthopaedics and related research.

[20]  B. Boyan,et al.  Pulsed electromagnetic field stimulation of MG63 osteoblast‐like cells affects differentiation and local factor production , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  A. Wolfman,et al.  Endogenous c-N-Ras Provides a Steady-state Anti-apoptotic Signal* , 2000, The Journal of Biological Chemistry.

[22]  B. Bianco,et al.  Zeeman-Stark modeling of the RF EMF interaction with ligand binding. , 2000, Bioelectromagnetics.

[23]  D. Alessi,et al.  Mammalian target of rapamycin is a direct target for protein kinase B: identification of a convergence point for opposing effects of insulin and amino-acid deficiency on protein translation. , 1999, The Biochemical journal.

[24]  Andrius Kazlauskas,et al.  Diverse Signaling Pathways Activated by Growth Factor Receptors Induce Broadly Overlapping, Rather Than Independent, Sets of Genes , 1999, Cell.

[25]  P. Krebsbach,et al.  Isolation and Characterization of MC3T3‐E1 Preosteoblast Subclones with Distinct In Vitro and In Vivo Differentiation/Mineralization Potential , 1999, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[26]  R. Doherty Long term follow up , 1999, BMJ.

[27]  C. Rubin,et al.  Effects of electromagnetic fields in experimental fracture repair. , 1998, Clinical orthopaedics and related research.

[28]  J. Avruch,et al.  Amino Acid Sufficiency and mTOR Regulate p70 S6 Kinase and eIF-4E BP1 through a Common Effector Mechanism* , 1998, The Journal of Biological Chemistry.

[29]  R. Pearson,et al.  Rapamycin suppresses 5′TOP mRNA translation through inhibition of p70s6k , 1997, The EMBO journal.

[30]  H. Liu,et al.  Pulsed electromagnetic fields influence hyaline cartilage extracellular matrix composition without affecting molecular structure. , 1996, Osteoarthritis and cartilage.

[31]  J. King,et al.  A prospective, double-blind trial of electrical capacitive coupling in the treatment of non-union of long bones. , 1994, The Journal of bone and joint surgery. American volume.

[32]  R. Bernstein,et al.  Treatment of ununited tibial fractures: a comparison of surgery and pulsed electromagnetic fields (PEMF). , 1992, Orthopedics.

[33]  D. Garland,et al.  Long-term follow-up of fracture nonunions treated with PEMFs. , 1991, Contemporary orthopaedics.

[34]  W. Sharrard,et al.  A double-blind trial of pulsed electromagnetic fields for delayed union of tibial fractures. , 1990, The Journal of bone and joint surgery. British volume.

[35]  R. Borgens Endogenous ionic currents traverse intact and damaged bone. , 1984, Science.

[36]  T. Martin,et al.  Morphological and biochemical characterization of four clonal osteogenic sarcoma cell lines of rat origin. , 1983, Cancer research.

[37]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[38]  M. Shamos,et al.  Piezoelectric Effect in Bone , 1963, Nature.

[39]  C. Andrew L. Bassett,et al.  Generation of Electric Potentials by Bone in Response to Mechanical Stress , 1962, Science.

[40]  Eiichi Fukada,et al.  On the Piezoelectric Effect of Bone , 1957 .

[41]  R. Aaron,et al.  Upregulation of basal TGFbeta1 levels by EMF coincident with chondrogenesis--implications for skeletal repair and tissue engineering. , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[42]  P. Strauss,et al.  Effects of extremely low frequency electromagnetic field (EMF) on collagen type I mRNA expression and extracellular matrix synthesis of human osteoblastic cells. , 1998, Bioelectromagnetics.

[43]  F. Marinelli,et al.  Intramembrane protein distribution in cell cultures is affected by 50 Hz pulsed magnetic fields. , 1997, Bioelectromagnetics.

[44]  C. Rubin,et al.  Electromagnetic fields in bone repair and adaptation , 1995 .

[45]  S. Pollack,et al.  The origin of stress‐generated potentials in fluid‐saturated bone , 1983, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.