cellular canceration induced by mobile phone.pdf

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Cellular Canceration Induced by Mobile Phone

RadiationYang Lei Wang MingLian Hao DongMei Zeng Yi

School of Life Science and Bioengineering

Beijing University of TechnologyBeijing, China

[email protected]

Abstract —With the rapid development of communicationtechnology, there is a widespread use of mobile telephone,whose effect on human has been getting more and moreattention. The electromagnetic wave can penetrate biosystemsand interacts with biological tissue at different biologicallevels. Although many researchers have been studying

biological effects of electromagnetic radiation for years, their results are not consistent and need to be confirmed by reliableand repeatable experiments. Since the energy of electromagnetic radiation of mobile phone is much low and theorganism has an adaptability to external stimulation, short-timeradiation may not induce obvious effect on organism. To avoidother inductive factors in the environment, we used in vitro cellculture technology and exposure experiment to study theactivating effect of 900 MHz electromagnetic fields (EMF) on

NIH/3T3 cells. NIH/3T3 cells are sensitive to sarcoma virusfocus formation and leukaemia virus propagation, and thereforeare often used for DNA transformation studies. The NIH/3T3cells were divided into four groups, the control group withsham exposure and the three exposure groups exposed to10W/m2, 50W/m2 and 90W/m2 916MHz EMF respectively.The exposure lasted 2 hours a day. After exposure we foundthe cells morphology changed and had the characteristics of

epithelial cells. The Soft agar assay showed that all theexposure groups formed colonies in soft agar while the controlgroup did not form colonies. The results in this study suggestthat 916MHz electromagnetic waves can promote malignanttransformation of NIH/3T3 cells and induce cellular canceration .

Keywords-mobile phone; Electromagnetic radiation; Biological

effect; cellular canceration

I. I NTRODUCTION

The rapid and more widespread use of mobile phones hascaused a great public concern regarding their possible

biological effects. Although no adverse health effects of mobile

phones have been confirmed scientifically, the data to assesstheir safety are still insufficient.

Many studies have investigated the biological effects of pulsed high frequency electromagnetic field (EMF) emitted bymobile phones with human and animal experiments. In theexperiments with human being, researchers recorded thesubjects’ physiological parameters such aselectroencephalograph (EEG), blood pressure, heart rate,

breath rate and regional cerebral blood flow et al. before andafter exposure to EMF to investigate the short-term effect of

mobile phones [1-4]. They also tested the hypothesis that long-term mobile phone use increases the risk of brain tumor [5].

In animal experiments, rats were usually used to expose toEMF with higher intensity for a longer time compared withhuman being to simulate the long-term exposure and then

behavior and physiological tests were done to indicate thechronic effects of EMF from mobile phones. Anne-LaureMausset-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 theamount of N-methyl-d-aspartate (NMDA) receptors at the

postsynaptic membrane. Margaret Tzaphlidou et al [7] reportedthat following 2 hours in vivo exposure to 900 MHz RF fieldsfor 30 days, the collagen fibril architecture was disturbed onlyin males while the fibril diameter was decreased in both malesand females. Thomas Tillmann et al [8] found no evidence thatthe exposure of male and female B6C3F1 mice to both 900MHz and high 1800 MHz GSM RF radiation at a whole-bodySAR of 4 W/kg induce any carcinogenic effect. Most studieson animal carcinogenicity found no evidence of carcinogeniceffects from RF field exposures. Several studies investigatedthe 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 fieldsinduce cell death [9-12]. Chronic and repeated experiments areneeded before getting clear conclusions. We hope our researchwill be helpful for further studies in this field.

This study was designed to investigate the cancer inductionof electromagnetic fields (EMF) emitted by mobile phone. Theshort-time high electromagnetic radiation was utilized tosimulate the long-time low mobile phone radiation. Weinvestigated cellular canceration by observing the NIH/3T3 cellsmorphological change.

II. MATERIALS AND METHODS

A. Materials cells culture

NIH/3T3 cells line was established from NIH Swissmouse embryo cultures in the same manner as the originalrandom bred 3T3 and the inbred BALB/c 3T3.

NIH/3T3 is an ideal experimental object in cellular canceration induced study. These fibroblasts cells process ahigh degree of contact inhibition and are sensitive to sarcomavirus focus formation and leukaemia virus propagation.

NIH/3T3 cells will lose their contact inhibition when they aretransformed, 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-mail：[email protected]

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



studies. Besides, they are immortalized cells which could becultured for more than 100 generations, and therefore aresuitable for long term experiments.

NIH/3T3 cells are adherent cells which grew in dulbecco'sminimum 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 theculture flask, cell passage should be taken in time otherwisecells would die due to over proliferation and exhaustednutrition in culture medium.

B. Electromagnetic field exposure system

The exposure system is composed of a 916MHzmicrowave 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 wavesimilar to a mobile phone. Its output power is adjustable withthe maximum 90W/m2 and the output frequency 916MHz. Aradiation meter produced by Chinese Radiation ResearchInstitute was used to measure the electromagnetic fielddistribution around the antenna. As we all know, the power at

one position in electromagnetic field is associated with itsdistance to the radiation source. Therefore different power could be obtained at different position in the EMF at the sametime. The power of 10W/m2, 50W/m2 and 90W/m2 were usedin our study, in which 10W/m2 approaches to the power of mobile phone.

Figure 1. Electromagnetic field exposure system

C. Exposure procedure

• NIH3T3 cells were divided into 4 groups. The controlgroup was not exposed to EMF. The three exposuregroups were located at the position of power 10W/m2,50W/m2 and 90W/m2 in the EMF for 2 hours everyday. The 10W/m2 group lasted for 8 weeks, 50W/m2group for 6 weeks and 90W/m2 group for 5 weeks

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

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

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 thestrictest 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 2×DMEM. Added 2ml of this mixture

to per well of the 6-well plate and set it aside to allowagar to solidify.

• Secondly, counted the number of cells. After preparation of single cell suspension, we advised tocount the number of cells per ml to ensure each grouphad the same concentration. It can be a help to ensurethe result was valid. This study required 100,000cells/ml and 0.2ml per well.

• Thirdly, prepared the top agar. Equal volume mixed

0.6% agar and 2×DMEM. Added 0.2ml single cellsuspension and 1ml the mixture to the well whichcontained the base agar. Placed this 6-well plate at

incubated box for 10-30 days.

III. RESULT

During exposure, apoptosis rate in 90W/m2 group washigher than the other groups. After 5 weeks, a large number cells in this group was detached and then stopped exposure. Aweek later, it was observed the 90W/m2 group cells lost their contact inhibition. In Fig.3, the cells morphology changed andhad the characteristics of epithelial cells. Although the cells in10W/m2 and 50W/m2 groups did not change their morphology



simultaneously, they changed their phenotypic after exposure 8weeks and 6 weeks respectively.

The transformation can be observed by certain phenotypicchanges such as loss of contact inhibition and anchorageindependence. The last one would be found by cells formingcolonies in soft agar. Soft agar assay was taken until all groupsstopped exposure. After 26 days, it was found that all threeexposure groups could form colonies in soft agar while thecontrol group was unable to form colonies. (see Fig.4).

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 atweek 5 and 6.

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 inhuman’s head might be cause by use mobile phone frequently.Therefore, the biological effect of electromagnetic wave on theorganism is of the highest concern to the public health. Yu et al[13] observed no evidence that GSM exposure of 900 MHz atSARs of 0.44, 1.33, or 4.0 W/kg promoted mammary tumor development in rats. However, Diem et al [14] found that low-level 1800-MHz RF field exposures induced primary DNAdamage 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 detectedat once. It is difficult to track long-time effect yet. Therefore,how to investigate the long term effect in short time is animportant problem in the study. We supposed that high power radiation in a short-time maybe possible to simulate the effectof low power in a long-time. We took two high power exposegroups 50W/m2 and 90W/m2 to study the biological effect of EMF and the relationship between the power and exposuretime.

The 90W/m2 group has a higher apoptosis rate than othersduring exposure. We found they changed their morphology andlost their contact inhibition after 5 weeks exposure and the

same changes were found in the other groups after 6~8 weekswhich indicated that those cells had transformed. Itdemonstrates the adverse effect of EMF on NIH/3T3 cells.

We took soft agar assay to further investigate these cellstransformation. The result is associated with the certain

phenotypic changes and shows that EMF induced all exposuregroups transformation. The soft agar result indicates therelationship between the power and exposure time, whichshows high electromagnetic radiation lasting a short time hasthe similar effect to low radiation lasting a long-time.

[email protected]



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.

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