Cellular Canceration Induced by Mobile Phone Radiation

Yang Lei Wang MingLian Hao DongMei Zeng Yi School of Life Science and Bioengineering Beijing University of Technology Beijing, China ylyjylyj@emails.bjut.edu.cn AbstractWith the rapid development of communication technology, there is a widespread use of mobile telephone, whose effect on human has been getting more and more attention. The electromagnetic wave can penetrate biosystems and interacts with biological tissue at different biological levels. Although many researchers have been studying biological effects of electromagnetic radiation for years, their results are not consistent and need to be confirmed by reliable and repeatable experiments. Since the energy of electromagnetic radiation of mobile phone is much low and the organism has an adaptability to external stimulation, short-time radiation may not induce obvious effect on organism. To avoid other inductive factors in the environment, we used in vitro cell culture technology and exposure experiment to study the activating effect of 900 MHz electromagnetic fields (EMF) on NIH/3T3 cells. NIH/3T3 cells are sensitive to sarcoma virus focus formation and leukaemia virus propagation, and therefore are often used for DNA transformation studies. The NIH/3T3 cells were divided into four groups, the control group with sham exposure and the three exposure groups exposed to 10W/m2, 50W/m2 and 90W/m2 916MHz EMF respectively. The exposure lasted 2 hours a day. After exposure we found the cells morphology changed and had the characteristics of epithelial cells. The Soft agar assay showed that all the exposure groups formed colonies in soft agar while the control group did not form colonies. The results in this study suggest that 916MHz electromagnetic waves can promote malignant transformation of NIH/3T3 cells and induce cellular canceration .
Keywords-mobile phone; Electromagnetic radiation; Biological effect; cellular canceration

mobile phones [1-4]. They also tested the hypothesis that longterm mobile phone use increases the risk of brain tumor [5]. In animal experiments, rats were usually used to expose to EMF with higher intensity for a longer time compared with human being to simulate the long-term exposure and then behavior and physiological tests were done to indicate the chronic effects of EMF from mobile phones. Anne-Laure Mausset-Bonnefont et al [6] observed neuronal damage in rat brain cells after in vivo exposure to 900MHz radio frequency (RF) fields for 15 minutes. They also observed decrease of the amount of N-methyl-d-aspartate (NMDA) receptors at the postsynaptic membrane. Margaret Tzaphlidou et al [7] reported that following 2 hours in vivo exposure to 900 MHz RF fields for 30 days, the collagen fibril architecture was disturbed only in males while the fibril diameter was decreased in both males and females. Thomas Tillmann et al [8] found no evidence that the exposure of male and female B6C3F1 mice to both 900 MHz and high 1800 MHz GSM RF radiation at a whole-body SAR of 4 W/kg induce any carcinogenic effect. Most studies on animal carcinogenicity found no evidence of carcinogenic effects from RF field exposures. Several studies investigated the relationship between RF radiation and the rate of cell proliferation and cell death. They found no effect of RF exposure on cell proliferation and no evidence that RF fields induce cell death [9-12]. Chronic and repeated experiments are needed before getting clear conclusions. We hope our research will be helpful for further studies in this field. This study was designed to investigate the cancer induction of electromagnetic fields (EMF) emitted by mobile phone. The short-time high electromagnetic radiation was utilized to simulate the long-time low mobile phone radiation. We investigated cellular canceration by observing the NIH/3T3 cells morphological change. II. MATERIALS AND METHODS

I.

INTRODUCTION

The rapid and more widespread use of mobile phones has caused a great public concern regarding their possible biological effects. Although no adverse health effects of mobile phones have been confirmed scientifically, the data to assess their safety are still insufficient. Many studies have investigated the biological effects of pulsed high frequency electromagnetic field (EMF) emitted by mobile phones with human and animal experiments. In the experiments with human being, researchers recorded the subjects physiological parameters such as electroencephalograph (EEG), blood pressure, heart rate, breath rate and regional cerebral blood flow et al. before and after exposure to EMF to investigate the short-term effect of

A. Materials cells culture NIH/3T3 cells line was established from NIH Swiss mouse embryo cultures in the same manner as the original random bred 3T3 and the inbred BALB/c 3T3. NIH/3T3 is an ideal experimental object in cellular canceration induced study. These fibroblasts cells process a high degree of contact inhibition and are sensitive to sarcoma virus focus formation and leukaemia virus propagation. NIH/3T3 cells will lose their contact inhibition when they are transformed, which are suitable for DNA transformation

This work was supported by grants from research projects of Beijing Municipal Education Commission (No. KM200910005016) Corresponding author: Hao dongmei, E-mailhaodongmei@bjut.edu.cn

978-1-4244-4713-8/10/$25.00 2010 IEEE

studies. Besides, they are immortalized cells which could be cultured for more than 100 generations, and therefore are suitable for long term experiments. NIH/3T3 cells are adherent cells which grew in dulbecco's minimum essential medium (DMEM) containing 5% fetal calf serum (FCS) and seeded in culture flask at incubated box. When monolayer cells formed and spread on the bottom of the culture flask, cell passage should be taken in time otherwise cells would die due to over proliferation and exhausted nutrition in culture medium. B. Electromagnetic field exposure system The exposure system is composed of a 916MHz microwave power generator and a monopole antenna offered by Tian Hua Zhong Wei Technology Co., Ltd., Beijing China (See Fig.1) The device can generate electromagnetic wave similar to a mobile phone. Its output power is adjustable with the maximum 90W/m2 and the output frequency 916MHz. A radiation meter produced by Chinese Radiation Research Institute was used to measure the electromagnetic field distribution around the antenna. As we all know, the power at one position in electromagnetic field is associated with its distance to the radiation source. Therefore different power could be obtained at different position in the EMF at the same time. The power of 10W/m2, 50W/m2 and 90W/m2 were used in our study, in which 10W/m2 approaches to the power of mobile phone.

respectively. (See Fig.2). The electromagnetic waves was 916MHz.

frequency

of

The morphologic features of cells were observed under inverted microscope each day and took pictures each week.

Figure 2. Position of exposure groups in the EMF

D. Soft agar assay for colony formation The soft agar assay for colony formation is an anchorage independent growth assay in soft agar. It is considered the strictest assay for verifying malignant transformation of cells. Two types of soft agar were used in this assay, which were base agar and top agar. Firstly, prepared the base agar. Equal volume mixed 1.2% agar and 2DMEM. Added 2ml of this mixture to per well of the 6-well plate and set it aside to allow agar to solidify. Secondly, counted the number of cells. After preparation of single cell suspension, we advised to count the number of cells per ml to ensure each group had the same concentration. It can be a help to ensure the result was valid. This study required 100,000 cells/ml and 0.2ml per well. Thirdly, prepared the top agar. Equal volume mixed 0.6% agar and 2DMEM. Added 0.2ml single cell suspension and 1ml the mixture to the well which contained the base agar. Placed this 6-well plate at incubated box for 10-30 days. III. RESULT

Figure 1. Electromagnetic field exposure system

C. Exposure procedure NIH3T3 cells were divided into 4 groups. The control group was not exposed to EMF. The three exposure groups were located at the position of power 10W/m2, 50W/m2 and 90W/m2 in the EMF for 2 hours every day. The 10W/m2 group lasted for 8 weeks, 50W/m2 group for 6 weeks and 90W/m2 group for 5 weeks

During exposure, apoptosis rate in 90W/m2 group was higher than the other groups. After 5 weeks, a large number cells in this group was detached and then stopped exposure. A week later, it was observed the 90W/m2 group cells lost their contact inhibition. In Fig.3, the cells morphology changed and had the characteristics of epithelial cells. Although the cells in 10W/m2 and 50W/m2 groups did not change their morphology

simultaneously, they changed their phenotypic after exposure 8 weeks and 6 weeks respectively. The transformation can be observed by certain phenotypic changes such as loss of contact inhibition and anchorage independence. The last one would be found by cells forming colonies in soft agar. Soft agar assay was taken until all groups stopped exposure. After 26 days, it was found that all three exposure groups could form colonies in soft agar while the control group was unable to form colonies. (see Fig.4).

Figure 4. The colony formation of 4 groups in soft agar. A: The control group; B: The 10W/m2 exposure group; C: The 50W/m2 exposure group; D: The 90W/m2 exposure group;

IV.

DISCUSSION

Several cases were reported that malignant tumor in humans head might be cause by use mobile phone frequently. Therefore, the biological effect of electromagnetic wave on the organism is of the highest concern to the public health. Yu et al [13] observed no evidence that GSM exposure of 900 MHz at SARs of 0.44, 1.33, or 4.0 W/kg promoted mammary tumor development in rats. However, Diem et al [14] found that lowlevel 1800-MHz RF field exposures induced primary DNA damage in a variety of mammalian cell lines. The majority of studies provided no evidence of genotoxic effects, but a few positive found were reported. The EMF have cumulate and delay effect, which means that short-time radiation may not produce any harmful influence or its effect can not be detected at once. It is difficult to track long-time effect yet. Therefore, how to investigate the long term effect in short time is an important problem in the study. We supposed that high power radiation in a short-time maybe possible to simulate the effect of low power in a long-time. We took two high power expose groups 50W/m2 and 90W/m2 to study the biological effect of EMF and the relationship between the power and exposure time. The 90W/m2 group has a higher apoptosis rate than others during exposure. We found they changed their morphology and lost their contact inhibition after 5 weeks exposure and the same changes were found in the other groups after 6~8 weeks which indicated that those cells had transformed. It demonstrates the adverse effect of EMF on NIH/3T3 cells. We took soft agar assay to further investigate these cells transformation. The result is associated with the certain phenotypic changes and shows that EMF induced all exposure groups transformation. The soft agar result indicates the relationship between the power and exposure time, which shows high electromagnetic radiation lasting a short time has the similar effect to low radiation lasting a long-time.

Figure 3. a,b:The control group at week 5 and 6; c, d 10W/m2 group at week 5 and 6,; e, f 50W/m2 group at week 5 and 6;, g, h 90W/m2 group at week 5 and 6.

Haodongmei@bjut.edu.cn

In summary, the results in this study suggest that high power 916MHz electromagnetic waves can induce cellular morphological change, which indicts canceration probability. In the next study, experimental inoculation of Scid mice would be done to validate the canceration.

V.
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