Changes in polyamines, c‐myc and c‐fos gene expression in osteoblast‐like cells exposed to pulsed electromagnetic fields

Pulsed electromagnetic field (PEMF) stimulation promotes the healing of fractures in humans, though its effect is little known. The processes of tissue repair include protein synthesis and cell differentiation. The polyamines (PA) are compounds playing a relevant role in both protein synthesis processes and cell differentiation through c‐myc and c‐fos gene activation. Since several studies have demonstrated that PEMF acts on embryonic bone cells, human osteoblast‐like cells and osteosarcoma TE‐85 cell line, in this study we analyzed the effect on cell PAs, proliferation, and c‐myc and c‐fos gene expression of MG‐63 human osteoblast‐like cell cultures exposed to a clinically useful PEMF. The cells were grown in medium with 0.5 or 10% fetal calf serum (FCS). c‐myc and c‐fos gene expressions were determined by RT‐PCR. Putrescine (PUT), spermidine (SPD), or spermine (SPM) levels were evaluated by HPLC. [3H]‐thymidine was added to cultures for DNA analysis. The PEMF increased [3H]‐thymidine incorporation (P ≤ .01), while PUT decreased after treatment (P ≤ .01); SPM and SPD were not significantly affected. c‐myc was activated after 1 h and downregulated thereafter, while c‐fos mRNA levels increased after 0.5 h and then decreased. PUT, SPD, SPM trends, and [3H]‐thymidine incorporation were significantly related to PEMF treatment. These results indicate that exposure to PEMF exerts biological effects on the intracellular PUT of MG‐63 cells and DNA synthesis, influencing the genes encoding c‐myc and c‐fos gene expression. These observations provide evidence that in vitro PEMF affects the mechanisms involved in cell proliferation and differentiation. Bioelectromagnetics 26:207–214, 2005. © 2005 Wiley‐Liss, Inc.

[1]  J. L. Phillips,et al.  Effect of 60 Hz magnetic field exposure on c-fos expression in stimulated PC12 cells , 1998, Molecular and Cellular Biochemistry.

[2]  Hiroyuki Aburatani,et al.  SB-431542 and Gleevec inhibit transforming growth factor-beta-induced proliferation of human osteosarcoma cells. , 2003, Cancer research.

[3]  S. Oredsson,et al.  Polyamine dependence of normal cell-cycle progression. , 2003, Biochemical Society transactions.

[4]  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.

[5]  Xiang-hang Luo,et al.  Effects of Progesterone and 18‐Methyl Levonorgestrel on Osteoblastic Cells , 2003, Endocrine research.

[6]  P. Diniz,et al.  Nitric oxide mediates the effects of pulsed electromagnetic field stimulation on the osteoblast proliferation and differentiation. , 2002, Nitric oxide : biology and chemistry.

[7]  P. Diniz,et al.  Effects of pulsed electromagnetic field (PEMF) stimulation on bone tissue like formation are dependent on the maturation stages of the osteoblasts , 2002, Bioelectromagnetics.

[8]  Ruggero Cadossi,et al.  Effect of low frequency electromagnetic fields on A2A adenosine receptors in human neutrophils , 2002, British journal of pharmacology.

[9]  S. Canaider,et al.  Epithelial-mesenchymal interactions and lung branching morphogenesis. Role of polyamines and transforming growth factor beta1. , 2009, European journal of histochemistry : EJH.

[10]  H Ito,et al.  The efficacy of ununited tibial fracture treatment using pulsing electromagnetic fields: relation to biological activity on nonunion bone ends. , 2001, Journal of Nippon Medical School = Nippon Ika Daigaku zasshi.

[11]  L. Wenger,et al.  Induction of Matrix Metalloproteinase-8 in Human Fibroblasts by Basic Calcium Phosphate and Calcium Pyrophosphate Dihydrate Crystals: Effect of Phosphocitrate , 2001, Connective tissue research.

[12]  F. Pezzetti,et al.  Effects of Pulsed Electromagnetic Fields on Human Articular Chondrocyte Proliferation , 2001, Connective tissue research.

[13]  H. Wiesmann,et al.  Capacitively coupled electric fields accelerate proliferation of osteoblast-like primary cells and increase bone extracellular matrix formation in vitro , 2000, European Biophysics Journal.

[14]  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.

[15]  S. Guggino,et al.  Thiazide Diuretics Affect Osteocalcin Production in Human Osteoblasts at the Transcription Level Without Affecting Vitamin D3 Receptors , 2000, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[16]  M. Maioli,et al.  Elf-pulsed magnetic fields modulate opioid peptide gene expression in myocardial cells. , 2000, Cardiovascular research.

[17]  Aaron,et al.  "Power frequency fields promote cell differentiation coincident with an increase in transforming growth factor-b(1) expression" , 1999, Bioelectromagnetics.

[18]  F. Pezzetti,et al.  Correlation between pulsed electromagnetic fields exposure time and cell proliferation increase in human osteosarcoma cell lines and human normal osteoblast cells in vitro. , 1999, Bioelectromagnetics.

[19]  J. Kanczler,et al.  Pulsed Electromagnetic Fields Simultaneously Induce Osteogenesis and Upregulate Transcription of Bone Morphogenetic Proteins 2 and 4 in Rat Osteoblastsin Vitro , 1998 .

[20]  M. Hüfner,et al.  In vitro differentiation potential of a new human osteosarcoma cell line (HOS 58). , 1998, Differentiation; research in biological diversity.

[21]  J. Risteli,et al.  Type I procollagen synthesis is regulated by steroids and related hormones in human osteosarcoma cells , 1998, Journal of cellular biochemistry.

[22]  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.

[23]  V. Kosma,et al.  Epidermal ornithine decarboxylase and polyamines in mice exposed to 50 Hz magnetic fields and UV radiation. , 1998, Bioelectromagnetics.

[24]  T. Hasuma,et al.  Growth-inhibitory effect of a high glucose concentration on osteoblast-like cells. , 1998, Bone.

[25]  Seymour S. Cohen A Guide to the Polyamines , 1998 .

[26]  K. H. Mild,et al.  ORNITHINE DECARBOXYLASE ACTIVITY AND POLYAMINE LEVELS ARE DIFFERENT IN JURKAT AND CEM-CM3 CELLS AFTER EXPOSURE TO A 50 HZ MAGNETIC FIELD , 1997 .

[27]  F. Pezzetti,et al.  Responses of human MG-63 osteosarcoma cell line and human osteoblast-like cells to pulsed electromagnetic fields. , 1997, Bioelectromagnetics.

[28]  D Krause,et al.  Role of modulation on the effect of microwaves on ornithine decarboxylase activity in L929 cells. , 1997, Bioelectromagnetics.

[29]  T. Litovitz,et al.  The role of temporal sensing in bioelectromagnetic effects. , 1997, Bioelectromagnetics.

[30]  F. P. Magee,et al.  Combined magnetic fields increase insulin-like growth factor-II in TE-85 human osteosarcoma bone cell cultures. , 1995, Endocrinology.

[31]  F. P. Magee,et al.  IGF‐II receptor number is increased in TE‐85 osteosarcoma cells by combined magnetic fields , 1995, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[32]  U. Bachrach,et al.  Activation of the proto-oncogene c-myc and c-fos by c-ras: involvement of polyamines. , 1994, Biochemical and biophysical research communications.

[33]  Gi Mammi,et al.  Effect of PEMF on the healing of human tibial osteotomies : a double blind study , 1993 .

[34]  D Krause,et al.  Effect of coherence time of the applied magnetic field on ornithine decarboxylase activity. , 1991, Biochemical and biophysical research communications.

[35]  B. Sacktor,et al.  Osteocalcin secretion by the human osteosarcoma cell line MG‐63 , 1990, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[36]  R. Goodman,et al.  Changes in levels of c-myc and histone H2B following exposure of cells to low-frequency sinusoidal electromagnetic fields: evidence for a window effect. , 1990, Bioelectromagnetics.

[37]  C. Baglioni,et al.  Tumor necrosis factor stimulates proliferation of human osteosarcoma cells and accumulation of c‐myc messenger RNA , 1988, Journal of cellular physiology.

[38]  W. R. Adey,et al.  The effects of low-energy 60-Hz environmental electromagnetic fields upon the growth-related enzyme ornithine decarboxylase. , 1987, Carcinogenesis.

[39]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[40]  C. Stefanelli,et al.  Separation of N1- and N8-acetylspermidine isomers by reversed-phase column liquid chromatography after derivatization with dansyl chloride. , 1986, Journal of chromatography.

[41]  C. Bassett,et al.  Pulsing electromagnetic field treatment in ununited fractures and failed arthrodeses. , 1982, JAMA.