General Scientific Researches, Vol(2), No (1), December, 2014. pp.

18-21

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General Scientific Researches

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The Influences of Electromagnetic Field Irradiated by High-Voltage

Transmission Lines with 50Hz on Properties of Cells
Pang Xiao feng *
Institute of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, China.

Chen Shude
Department of Physics, East China Normal University, Shanghai, 200062, China.

Zhong Lisheng
State Key Lab. of Electrical Insulation and Power Equipment, Xian Jiaotong University, Xian 710049, China.
*Corresponding author: pangxf2006@aliyun.com

Keywords

Abstract

Electromagnetic radiation,
High-voltage transmission lines,
Biological effect,
Signal transduction,
Cell

The influences of low-frequency electromagnetic field irradiating by high-voltage transmission lines on

signal transduction of cell in spleen cells of the rates have been studied by molecular-biology techniques.
The spleen cells are extracted from skilled rats, which are exposed in the electromagnetic field of highvoltage transmission lines with 4000V/m and 0.09-0.1G and 50Hz about 400days. The quantity or level of
Phosphorylation of signal transducer and activator of transcription (STAT3) in JAK-STAT signal
transduction pathway of spleen cells, which are stimulated and unstimulated by IL-2, respectively, are
detected by the immunoblotting and immunobiochemistry. The results show that the expression of phosphoSTAT3 in spleen cell stimulating by IL-2 differ not from that in the unstimulated cell. The former is
significantly large than the latter. This shows that signal transduction of cell is affected by this
electromagnetic field.

1.

Introduction

As known, the problem on influences of the electromagnetic fields (EMF) with 50 Hz irradiated by high-voltage transmission lines on the
persons health have been always considered and studied by scientists. Epidemiological investigations show that the EMF of high- voltage
transmission lines is always harmful to the health of men and women. However, the biological effects have not been systematically and
completely studied up to now, although a large number of reports on this problem have been published in a lot of journals, in which the results
obtained by different group and persons are different. Thus this problem is disputed long times [1-14]. In such a case it is very necessary to study
seriously and clarify its biological effects. We recently studied completely the influences of the EMF of the high-voltage transmission lines on
procreate organs, blood features, learing and memiry, signal transduction and prolifiration of cells in the rats by high-through techniques. The
results obtained show that the influences of the EMF on these tissues are larger. We here report the influence of the high-voltage transmission
lines on signal transduction of cells in the rats.
So-called signal transduction is just the process of transport of extracellular signals into interior of cell through membrane. The signals imported
can regulate gene expression and facilitate the cellular activation, proliferation, differentiation and damping of the cells. Generally speaking,
there are three pathways to transport the signals in the living system, the Janus kinase (JAK)-signal transducer and activator of transcription
(STAT) pathway [15-20] is one of three pathways. The JAKs associated with the membrane proximal region of a cytokine receptor can be
activated via ligand-induced dimerization of the receptor chains. Activated JAK proteins phosphorylate the receptor on a number of tyrosine
residueswhich creates the docking sites for proteins containing the SH2-domain. The STAT protein binds often to one of the phosphorylated
tyrosine residues on the receptor, and is also phosphorylated on the tyrosine residues by the activated JAKs. Phosphorylated STATs
homodimerize or heterodimerize each otherand translocate again to the nucleus where they can activate the transcription of target genes [18].
The JAK kinases are required for tyrosine phosphorylation and activation of latent cytoplasmic transcription factors. As known, the tyrosine
phosphorylation of the STATs is essential for activation, dimerizationand DNA binding [19]. The STAT3 in the STAT family is involved into
the process of the JAK/STAT signal transduction stimulated by IL-2. The activated STAT3 plays important roles in preventing apoptosis [16],
mediating cell cycle progression [17] and promoting tumor cell development and/or growth [15]. In this paper, by using Immunoprecipitation
and Western blot techniques, we analyze phosphorylation of STAT3, which activates the JAK-STAT Pathway, Since this pathway is easily
stimulated and activated by numerous electromagnetic fields, cytokines and growth factors. Therefore we here study influences of the EMF
irradiated by high-voltage transmission lines on the signal transduction of cells in the spleen cells through this pathway by immunoprecipitation
and immunoblotting ways.

2.

Methodology

In this experiment the materials used have rabbit anti-STAT3 polyclonal antibodies which is purchased from BOSHIDE biotechnology
Company, Protein G agarose which is purchased from SIGMA biotech, anti-phosphor-stat3 (Tyr705) antibodies which is purchased from Santa
Cruz biotech, anti-IgG mAb and horseradish peroxidase streptavidin which is purchased from Bei Jing Zhongshan biotechnology Company.

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The Influences of Electromagnetic Field Irradiated by High-Voltage Transmission Lines with 50Hz on Properties of Cells
General Scientific Researches Vol(2), No (1), December, 2014.

The 32 Wistar rats in 4th weeks were divided into controlled and experimental groups. The experimental group is exposed in this
electromagnetic field about 400days. An electromagnetic field of high-voltage transmission lines was generated by an appliance. The strengths
of the electric and magnetic fields for this electromagnetic field used are 4000V/m and 0.09-0.1G, respectively, which are the strengths of the
electric and magnetic fields at the position of 2.0- 2.3 m distancing the earth under the high-voltage transmission lines of 200KV, which was
concretely measured and obtained by us in surrounding of the high-voltage transmission lines of 220KV in Beijing and Chengdu. The spleen
cells used are extracted from the spleen of skilled rates after 400days exposed. The cells are scraped into ice-cold PBS and collected and washed
about twice by ice-cold PBS. After this, 4106 cells are resuspended in pellet, which are again divided into two parts, one of which was
stimulated with IL-2 (500U/ml), other is not. For preparation of lysates for immunoprecipitation 2106 cells /experiment were lysed in 100L
lysis buffer (50mM Tris PH7.4, 150 mM NaCl, 1mM EDTA, 1%NP-40, 1mM NaF and NaVO3 and the following inhibitors: 1mM ZnCl2,
0.1mM PMSF20mg/L Aprotinin) for 30min on ice. After spinning the samples for 10 min at 12 000 rpm, the supernatants are utilized for
Western blot analysis. In immunoprecipitation experiment, the anti-Stat3 antibodies are precleared for 2h with protein-G beads before incubation
with cell lysates overnight. Complexes are recovered by incubation with protein-G beads for 24h, followed by repeated washing of the
precipitates in lysis buffer. Samples are boiled in reducing SDS-sample buffer, separated on an 8% SDS-polyacrylamide gel electrophoresis(SDS-PAGE), and again transferred electrophoretically to a nitrocellulose membrane. The membrane is blocked in 5%BSA in PBS for
30min and incubated by anti-phospho- Stat3(Tyr7O5) antibodies in blocking buffer, followed by washing in PBS and incubation with
Biotinylated anti-IgG and Horseradish Peroxidase Streptavidin. Quantitative analysis of Western blot was performed by using a computerized
digital image analysis system (ShangHai FURI technology Company, ShangHai,CHINA).

3.

Experimental Results

We first measure the phosphor-STAT3 expression in spleen cells in the two groups, unstimu- lating by IL-2 by using western blotting of the
Immunoprecipitation. The result is shown in Fig.1, which is obtained by the following way. Cytoplasmic lysates in spleen cells unstimulated by
IL-2 from the exposed group (lanes 1, 2, 3, 4) and control group (lanes 5, 6, 7, 8) are subjected to SDS-PAGE and immunoblotted by antiphosphor-Stat3 (Tyr705) antibodies. From this figure we see that there are not remarkable difference in phosphor-STAT3 expression in spleen
cells for the controls and the exposed groups, when unstimulated with IL-2.
We measure further the phosphor-STAT3 expression in spleen cells in the two groups, stimulating by IL-2 (we know that the IL-2 can active the
JAK-STAT signal transduction pathway). We find that the phosphor-STAT3 expression increased dramatically both of two group samples after
stimulation of IL-2. Thus the up-regulation of phosphor-STAT3 expression is more remarkable in spleen cells in the exposed groups, when
compared with that of the controls, as shown in Fig.2. The way getting this result is as follows. The cells first are incubated by IL-2 in the
determinant time. The cells are next lysed, the cytoplasmic extracts in the cells are further immunoprecipitated by anti-Stat3 mAb from the
exposed group (lanes 1, 2, 3, 4) and the control group (lanes 5, 6, 7, 8). The precipitated proteins used are subjected to SDS-PAGE and
immunoblotted by anti-phosphor- Stat3 (Tyr705) antibodies.
In this experiment the western blotting data for the relative values of phosphor-STAT3 obtained by using a computerized digital image analysis
system are shown in Fig.3. From this figure we see that the increment of the western blotting data for the phosphor-STAT3 expression in the
spleen cells are not remarkable between the controlled and the exposed groups, when unstimulated with IL-2, but quite remarkable between the
exposed and controlled groups, when stimulated with IL-2.

Fig.1. Phosphor-STAT3 expression in spleen cells of rat unstimulated with IL-2. Here,
1- 4 denote phosphor-STAT3 expressions in the lanes 5 - 8 for the control group, respectively.

Pang Xiaofeng *, Chen Shude , Zhong Lisheng

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General Scientific Researches Vol(2), No (1), December, 2014.

Fig.2. Phosphor-STAT3 were expressed in rat spleen cells stimulated with IL-2. Here, 1- 4 denote the phosphor-STAT3 expressions in the
lanes 1- 4 for the exposed group, respectively; 5- 8 show the phosphor-STAT3 expressions in the lanes 5 - 8 for the control group, respectively.

To besides the possibility that the augmentation of phosphor-STAT3 expression is due to the stimulation of IL-2, we analyzed significant
difference of the extent of up-regulation of phosphor-STAT3 expression (Fig.3). These results clearly demonstrated that phosphor-STAT3
expression was augmented by the costimulation of IL-2 and electromagnetic field in spleen cells, and the electromagnetic field produced by
high-voltage transmission lines influenced phosphor-STAT3 expression in cells upon stimulation of IL-2.

Fig.3. Comparison on western blotting data for relative values of phosphor-STAT3. The Western blotting data are estimated using a computerized digital image
analysis system. Here, 1(the control group) and 2 (the exposed group) are relative values of phosphor-STAT3 in the spleen cells unstimulated with IL-2,
respectively; 3 (the control group) and 4 (the exposed group) are relative values of phosphor-STAT3 in the spleen cells stimulated with IL-2, respectively.

4.

Conclusion

We here choose the STAT3 activation as the parameter to express the influence of the electromagnetic field of high-voltage transmission lines
on the signal transduction of cell in the JAK-STAT pathway. The western blot analysis reveals clearly that the phosphor-Stat3 expression was
remarkably upregulated in spleen cells stimulated simultaneously by IL-2 and the electromagnetic field of High-Voltage transmission lines.
Whereas the phosphor-Stat3 expression was slightly upregulated in these of the spleen cells stimulated only by IL-2 in the control group. This
denotes clearly that the electromagnetic fields of high-voltage transmission lines accelerate the process of phosphorylation of STAT3 activated
by IL-2, thus it activate also the JAK-STAT signal transduction pathway. Since the JAK/STAT signal transduction pathway play crucial roles in
regulating gene expression and cellular activation, then, we can infer that the electromagnetic field of high-voltage transmission lines can result
in a lot of biological effects of the cells and proteins in the activated JAK-STAT pathway in the living system.

Acknowledgements
The authors would like to acknowledge the National 973project of China for financial support (grate No: 212011CB503701).

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The Influences of Electromagnetic Field Irradiated by High-Voltage Transmission Lines with 50Hz on Properties of Cells
General Scientific Researches Vol(2), No (1), December, 2014.

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