Radio frequency radiation, small risk for some but very significant hazard for others, the p53 connection. By Dr Chris Barnes, Bangor Scientific and Educational Consultants, e-mail email@example.com
Dr Barnes Homepage http://www.drchrisbarnes.co.uk
Thousands of studies mainly on in vivo animal and human cell lines and some animal studies produce mixed and inconclusive results. Of the far fewer number of epidemiological studies, most suggest small but significant risks. Recently, however, Barnes (2013 (3 papers) ) considering meta-analysis for known UK and World Cancer incidence versus the growth of TV licensing in the UK and the growth of mobile wireless technologies worldwide has shown that such technologies produce far more significant risk and indeed in some Welsh housing estates the RF field levels seem consistently to be above average close to those properties of newly diagnosed cancer victims. By considering RF interactions with the p53 system and mutations of the system for specific cancers, the conclusion is reached that Radio frequency exposure may cause serious problems for a subset of the community with genetic deficiencies in the p53 tumour suppressor gene. Interaction of RF in general might be with a relatively small number of genes. Mechanisms may be by up regulation in Heat Shock Proteins or by more indirect routes. A number of cancers have been identified which are more susceptible to RF promotion. The present author has shown elsewhere that RF may also act as a co-promoter with ionising radiation and with other known carcinogens. Until such time as the precise mechanisms of chromosomal interaction are known and genetic testing can be put in place it would be prudent for everyone to take more precautions with RF exposure. With the exception of myelomas and melanomas vitamin D3 and Melatonin may conceivably offer some protection against 'RF' cancers. Natural herbal compounds like Milk Thistle may be useful for a number of cancers including Melanoma.
There has been growing concern regarding the safety of RF technology devices such as mobile phones, DECT phones and WIFI.
Thousands of studies mainly on in vivo animal and human cell lines and some animal studies produce mixed and inconclusive results. Of the far fewer number of epidemiological studies, most suggest small but significant risks.
Recently, however, I have shown by geographic meta-analysis for known UK and World Cancer incidence a strong correlation with the growth of TV licensing in the UK and with the growth of mobile wireless technologies worldwide. When the relevant statistics are applied this tends to suggest that the use of such technologies may produce higher relative risks (RR) than previously observed. I have also noted a much raised risk in some Welsh housing estates, wherein when the measured RF field levels seem consistently to be above average such locations seem to be at or close to those properties of newly diagnosed cancer victims which have been reported to me. The sample size here is I confess inevitably small.
I pose the question how can we rationally reconcile this? A probable and common sense hypothesis is to suggest that the genetic background may determine cellular responses to RF radiation. Thus of particular interest is p53, the tumour suppression gene.
This paper explores the evidence for this hypothesis.
Previous publications on RF suggestive of the hypothesis
Czyz et al (2004)  have used electromagnetic fields (EMF) simulating exposure to the Global System for Mobile Communications (GSM) signals on pluripotent embryonic stem (ES) cells in vitro. Wild-type ES cells and ES cells deficient for the tumor suppressor p53 were exposed to pulse modulated EMF at 1.71 GHz, lower end of the uplink band of GSM 1800, under standardized and controlled conditions, and transcripts of regulatory genes were analyzed during in vitro differentiation. Two dominant GSM modulation schemes (GSM-217 and GSM-Talk), which generate temporal changes between GSM-Basic (active during talking phases) and GSM-DTX (active during listening phases thus simulating a typical conversation), were applied to the cells at and below the basic safety limits for local exposures as defined for the general public by the International Commission on Nonionizing Radiation Protection (ICNIRP). They found that GSM-217 EMF induced a significant up regulation of mRNA levels of the heat shock protein, hsp70 of p53-deficient ES cells differentiating in vitro, paralleled by a low and transient increase of c-jun, c-myc, and p21 levels in p53-deficient, but not in wild-type cells. No responses were observed in either cell type after EMF exposure to GSM-Talk applied at similar slot-averaged specific absorption rates (SAR), but at lower time-averaged SAR values. Cardiac differentiation and cell cycle characteristics were not affected in embryonic stem and embryonic carcinoma cells after exposure to GSM-217 EMF signals. Thus their data indicate that genetic background determines cellular responses to GSM modulated EMF.
Working with yeast cells as a model, Chen at al 2012  produced results suggesting that the yeast cells did not alter gene expression in response to 50 Hz ELF-MF and that the response to RF-EMF is limited to only a very small number of genes, see Bioelectromagnetics 33:550–560, 2012. This is again in support for the hypothesis put forward in this present work.
Besides well-known dependencies on frequency and modulation, Belyaev (2005)  has reviewed and produced data suggesting dependencies of non-thermal microwave effects on intermittence and coherence time of exposure, polarization, genotype, gender, physiological and individual factors, static magnetic field, electromagnetic stray field, cell density during of exposure, and indicate that duration of exposure may be more important than power density for such effects. Again genotype was considered important, see http://informahealthcare.com/doi/abs/10.1080/15368370500381844
There is considerable disagreement as to whether RF can actually cause genetic aberrations (genotoxic effects) but it does seem to act as a promoter enhancing the probability of cancer in animal systems already carrying an oncogene, see for instance Repacholi et al (1997) . They looked at mice and concluded long-term intermittent exposure to RF fields can enhance the probability that mice carrying a lymphomagenic oncogene will develop lymphomas. Interestingly there is a study looking at p53 mutations in Merseyside suggesting that other carcinogens in addition to those in tobacco smoke may be involved in NSCLC in the Merseyside area of the UK, see Liloglou 1997  , possibly asbestos is the culprit, see Liu et al (1998) .
In summary RF might only cause problems in p53 deficient cells i.e. for a specific subset of the community. This might also explain RF 'sensitivity'.
Other RF interaction mechanisms and the p53 connection
EMFs disturb immune function through stimulation of various allergic and inflammatory responses, as well as effects on tissue repair processes, see Johansson (2009) . ROS and NOS has been associated with both allergic response, see Okayama (2005) and RF exposure, cited at . I have provided models as to how RF could mediate ROS on my website . The link between Inflammation and Cancer has been discussed by Schetter (2009) who discussed microRNA, free radical, cytokine and p53 pathways . Ö Hallberg, O Johansson (2011)  have suggested that melanoma incidence in Sweden has increased not because of any more DNA damage but due to problems with the body’s repair system brought on by FM broadcasting interfering with the immune system.
The ionising radiation connection.
I have produced epidemiological evidence suggesting that that 'RF' act as a co- promoter with ionising radiation or with atmospheric pollution . Properly controlled this might open up new possibilities in cancer treatment as well as a better understating of carcinogenic processes. For instance it is known that p53-dependent apoptosis modulates the cytotoxicity of anticancer agents, see Lowe et al 1993 . Loss of p53 function can make cancer harder to treat as evidenced, for instance by triple negative breast cancer. In tumours from radon exposed lung cancer patients, the mutation produced an amino acid change and an increased nuclear content of p53 protein, see Vahakangas et al (1992) .
In a complex organism, somatic cells are under intermittent selection pressure for the emergence of mutants that can survive environmental insults and that can grow autonomously despite adverse conditions. Repeated rounds of mutation, selection, and proliferation may lead to cancer. The organism prevents malignant transformation by assuring accurate DNA repair before cell division, by forcing the death of cells with excessive DNA damage, and by placing limits on the replicative life spans of most somatic cells. The p53 gene is a "guardian of the genome"—it regulates multiple components of the DNA damage control response and promotes cellular senescence. Disabling mutations and deletions of p53 occur in 50% of human tumours. P53-deficient cancers are often unstable, aggressive, and resistant to therapy.
Inactivation of p53, the tumour suppressor gene, contributes to the genesis and/or progression of a substantial fraction of all human cancers, including ≥50% of breast, lung, and colon carcinomas. I have shown the following specific cancers to have a strong ‘RF’ association. They are; Bowel, Brain, Breast, Kidney, Leukaemia, Malignant Melanoma, Multiple Myeloma and Prostate Cancer. These cancers will next be discussed with regard to the p53 gene.
Overall, Bosari et al (1995)  showed 89% of bowel carcinomas displayed p53 gene mutations and/or p53 accumulations of any type.
p53 mutation is also associated with Brain tumour progression, see Sidransky et al (1992)  . Further, mutations of the p53 gene have been found in conjunction with chromosome 17p, see Mashiyama et al (1991).
I have showed the strongest association of RF with breast cancer . It is thus common sense to explore the link with the p53 gene. Bartek et al (1990) considered the expression of the tumour suppressor gene p53 which was analysed in 11 human breast cancer cell lines by immunohistochemistry, immunoprecipitation and cDNA sequencing. They used a panel of anti-p53 monoclonal antibodies for cell staining and found abnormalities in every case. Eight of the cell lines produced a form of p53 which could be immunoprecipitated by the monoclonal antibody PAb240 but not by PAb1620. In the murine system PAb240 only immunoprecipitates mutant p53. They sequenced p53 cDNA directly from four of the PAb240 positive cell lines using asymmetric PCR templates. All four contained missense mutations in p53 RNA, with no detectable expression of the wild type sequence. Different residues were affected in each cell line, but all the mutations changed amino acids conserved from man to Xenopus. Their results imply that as in the murine system, the PAb240 antibody reliably detects a wide variety of p53 mutations and that these mutations have a common effect on the structure of p53. See CRC Handbook of Gene Level Diagnostics in Clinical Practice
By Victor A. Bernstam. .
Thus immunohistochemical data suggest that p53 mutation is the commonest genetic alteration so far detected in primary breast cancer, particularly aggressive 'triplenegative' breast cancers.
Thus it would appear we now may have the crucial link between RF exposure and breast cancer.
Reiter et al (1993)  suggests that, while the primary disease gene for kidney cancer appears to be on chromosome 3, abnormalities of p53 are common and may be involved in the progression of this malignancy. I would wish to further point out that in common with Brain Cancer above there may be Chromosome 17p Deletions and p53 Mutations in Renal Cell Carcinoma.
P53 alterations have also been noted in Leukaemia, see Ahuja et al 1989 . Fenaux et al (1991) suggest that alterations of the P53 gene may have a role in leukemogenesis in some cases of AML . The fact that P53 gene mutations occurred more often in patients with 17p monosomy seemed to support the “recessive” model of tumour suppressive activity of the P53 gene rather than the “dominant” model, in which alteration of only one allele is sufficient for the development of malignancy.
Stretch et al (1991)  showed 85% of specimens from a range of primary and metastatic melanomas had detectable evidence of p53 gene mutation, by virtue of the immunohistochemical detection of mutant p53 protein. Significantly increased prevalence of mutant p53 was found in metastatic melanoma, compared with primary tumors (P < 0.05). Their findings represent one of the highest incidences of this oncogenic mutation yet recorded in a human malignancy and support the concept that p53 may have a functional role in development of the metastatic tumor phenotype.
In Myeloma on the other hand P53 deletions are quite rare events, see Preudhomme 1992 with about 4% incidence only but other studies suggest there is still involvement of the short arm of chromosome 17 and also involvement of chromosome 6 and 13.
The data of Brookstein et al (1993)  indicate that mutated p53 alleles are quite uncommon in early prostate cancers but are found in 20–25% of advanced cancers, suggesting a role for p53 mutation in the progression of at least a subset of prostate cancers.
The melatonin connection
Of all 8 ‘RF’ cancers which I have previously studied , the incidence of 6 showed a significant correlation with night-time light pollution perhaps indicative of melatonin deficiency. Others have suggested that RF energy and/or ELF magnetic fields have a direct effect on the brain, hence suppressing melatonin. Developed areas of the world with a lot of night-time lighting also tend be the ones with a lot of wireless penetration. Further in that study, only melanoma and myeloma showed little or weak correlation with both melatonin and vitamin D.
Possibly the crucial link of melatonin with the above and with RF damage is in its potential modulation of cell-cycle length through control of the p53–p21 pathway, see Schernhammer, and Schulmeister 2004 .
Observational studies support an association between night work and cancer risk. We hypothesise that the potential primary culprit for this observed association is the lack of melatonin, a cancer-protective agent whose production is severely diminished in people exposed to light at night, see Schernhammer and Schulmeister (2004) .
Clinically, melatonin has a potential for a prevention or treatment of colorectal cancer, ulcerative colitis, irritable bowel syndrome, children’s colic and diarrhoea, see Bubenik (2001).
Pre-treatment of rats with intraperitoneal doses of 100 mg/kg melatonin provided a significant decrease in the DNA strand breakage and lipid peroxidation in rat brains as a result of irradiation, see Ündeğer et al (2004) in Melatonin: From Molecules to Therapy
edited by S. R. Pandi-Perumal, Daniel P. Cardinali .
Melatonin at physiological concentrations caused decreased MCF-7 human breast cancer cell decreased cell proliferation was coincident with a significant increase in the expression of p53 as well as p21WAF1 proteins. Melatonin appears to inhibit MCF-7 cell proliferation by inducing an arrest of cell cycle dependent on an increased expression of p21WAF1 protein, which is mediated by the p53 pathway, see Molis et al 2013 .
Possibly, crucially, Otkem et al (2005)  have shown that melatonin may exhibit a protective effect on mobile phone-induced renal impairment in rats. Hopefully, thus, Melatonin could be used to protect humans.
Kubo et al (2006)  have shown that compared with day workers, rotating-shift workers were significantly at risk for prostate cancer (relative risk = 3.0, 95% confidence interval: 1.2, 7.7), whereas fixed-night work was associated with a small and non- significant increase in risk. Their report was the first known to reveal a significant relation between rotating-shift work and prostate cancer and presumably has a link with serum melatonin across the entire circadian cycle .
The Vitamin D link
p53 is the most frequently mutated tumor suppressor gene in human neoplasia and encodes a transcriptional coactivator. Identification of p53 target genes is therefore key to understanding the role of p53 in tumorigenesis. To identify novel p53 target genes, Maruyama et al (2006)  first used a comparative genomics approach to identify p53 binding sequences conserved in the human and mouse genome. They hypothesized that potential p53 binding sequences that are conserved are more likely to be functional. Using stringent filtering procedures, 32 genes were newly identified as putative p53 targets, and their responsiveness to p53 in human cancer cells was confirmed by reverse transcription-PCR and real-time PCR. Among them, they focused on the vitamin D receptor (VDR) gene because vitamin D3 has recently been used for chemoprevention of human tumours.
VDR is induced by p53 as well as several other p53 family members, and analysis of chromatin immunoprecipitation showed that p53 protein binds to conserved intronic sequences of the VDR gene in vivo. Introduction of VDR into cells resulted in induction of several genes known to be p53 targets and suppression of colorectal cancer cell growth. In addition, p53 induced VDR target genes in a vitamin D3-dependent manner. Their in- silico approach is a powerful method for identification of functional p53 binding sites and p53 target genes that are conserved among humans and other organisms and for further understanding the function of p53 in tumorigenesis, see  Cancer Res 2006; 66(9): 4574-83).
Garland (1980)  proposed that vitamin D is a protective factor against colon cancer. This hypothesis arose from inspection of the geographic distribution of colon cancer deaths in the U.S., which revealed that colon cancer mortality rates were highest in places where populations were exposed to the least amounts of natural light - major cities, and rural areas in high latitudes. The hypothesis is supported by a comparison of colon cancer mortality rates in areas that vary in mean daily solar radiation penetrating the atmosphere. A mechanism involving cholecalciferol (vitamin D3) is suggested. This has recently been supported by my World Study of Barnes and by my USA study both found at reference .
Intake of 2000 IU/day of Vitamin D3, and, when possible, very moderate exposure to sunlight, could raise serum 25(OH)D to 52 ng/ml, a level associated with reduction by 50% in incidence of breast cancer, according to observational studies, see Garland 2007 . Again I have epidemiologic findings which show strong support for this .
Since breast cancer is also the most strongly correlated ‘RF’ cancer, vitamin D3 may hold out promise as a prophylactic here also?
Other natural compounds which may help
There is much interest in natural compounds as alternative and/or adjunct cancer therapies. It is proposed here that natural compounds which either reduce oxidative stress, boost the immune system or modulate cell survival may also be effective in reducing 'RF' promotion of cancer, especially if they modulate p53.
One such compound is silibinin found in the natural herbal plant milk thistle. Silibinin modulates mitogenic and survival signalling, p53, Cip1/p21 and other cell cycle regulatory molecules to prevent UVB-induced skin carcinogenesis. Studies also suggest the positive effect of silibinin on the repair of UVB-induced DNA damage in mouse skin. Overall, the protective efficacy of silibinin against skin cancer is supported by sound mechanistic rationale in animal and cell culture studies, and suggests its potential use for humans, see Singh and Agarwal (2005) .
Silymarin, a flavonoid antioxidant isolated from milk thistle, exerts exceptionally high to complete anticarcinogenic effects in tumorigenesis models of epithelial origin and in certain breast cancer cell lines (see Xi et al 1998). Silymarin induces apoptosis primarily through a p53-dependent pathway involving Bcl-2/Bax, cytochrome c release, and caspase activation, see Katiyar et al (2005) and Karimi .
Radio frequency exposure may potentially cause serious problems for a subset of the community with genetic deficiencies in the p53 tumour suppressor gene or to those exposed to ionising radiation or other carcinogens which may cause mutations or delations. Interaction of RF in general might be with a relatively small number of genes. Mechanisms may be by up regulation in Heat Shock Proteins or by more indirect routes, such as ROS and NOS and disturbance of Calcium channel pathways. A number of cancers have been identified which are more susceptible to RF promotion. These are cancers associated with oncogenes where there are p53 inactivation, deletions or aberrations. I have shown elsewhere that RF may also act as a co-promoter with ionising radiation and with other known carcinogens. Of course ionising radiation can cause genetic mutation of p53 and the like even in cases where there was initially no aberration, see Vahakangas et al (1992) .
Until such time as the precise mechanisms of chromosomal interaction are known and genetic testing can be put in place it would be prudent for everyone to take more precautions with RF exposure.
With the exception of myelomas and melanomas vitamin D3 and Melatonin may conceivably offer some protection against 'RF' cancers. Care should be taken with heavy Vitamin D supplementation because serum D3 and melatonin are negatively correlated, see Golan et al (2013) . This may be another way in which communities like the Amish get their apparent cancer protection. Living mainly in Pennsylvania, Ohio and Indiana they will get the full benefit of Vitamin D working out in the fields in summer but also the full benefit of melatonin by early to bed dark winter nights and only dim candle or oil lighting before retirement. In modern society we often lack both. I have commented on the Amish elsewhere with regard to EMF and RF .
Natural herbal compounds like Milk Thistle may be useful for a number of cancers including Melanoma.
It is imperative that cross disciplinary efforts between geneticists, molecular biologists and radio engineers are now multiplied to find cure or prevention for ‘RF’ promoted cancers a small risk for some but very significant hazard for others of a particular genetic makeup.
Perhaps one day soon there will be genetic testing and even gene therapy or targeted remediation for all oncogene type cancers, ‘RF’ promoted types included, see Felsher (2004) .
1. Bioelectromagnetics 25:296–307, 2004. http://www.ncbi.nlm.nih.gov/pubmed/15114639
16. CRC Handbook of Gene Level Diagnostics in Clinical Practice By Victor A. Bernstam.
23. From Molecules to Therapy edited by S. R. Pandi-Perumal, Daniel P. Cardinali.
27. Comparative Genome Analysis Identifies the Vitamin D Receptor Gene as a Direct Target of p53-Mediated Transcriptional Activation, Cancer Res 2006; 66(9): 4574-83).http://cancerres.aacrjournals.org/content/66/9/4574.full
32. Vahakangas, K.H., J.M. Samet, R.A. Metcalf et al. Mutations of p53 and ras genes in ... Lancet 339(8793): 576-580 (1992).