Molecular Screening for P 53 Mutations among Tobacco Smokers in a Surveyof Awareness of Links between Tobacco , Alcohol Use and Cancer in Saudi Arabia

Tobacco and alcohol consumption play an important role in mortality and morbidity in various countries (Ahmed, et al., 2003; Lopez, et al., 2006; Begg et al., 2007; Ahmed, et al., 2010). Globally, tobacco consumption is the main cause of death, resulting in millions of deaths every year, more than HIV/AIDS, tuberculosis and malaria (WHO, 2008). Cigarette and Shisha smoking are important public health concern worldwide causing many diseases including; pulmonary disease, various cancers including those of the respiratory, digestive, and genitourinary systems, some leukemias and premature death (Office of the Surgeon General, 2004; Babiker et al., 2012). Socioeconomic factors have strong impact on tobacco use worldwide (Bobak et al., 2000; Ahmed and Mahgoob, 2007). A strong inverse relationship between socioeconomic status and smoking exists, particularly in the develop world. Such relationship indicating that the poorest and least educated populations are more likely to smoke than the other counterparts (Laaksonen et al., 2005; Wipfli and Samet, 2009). Alcohol consumption is associated with high mortality and morbidity in both developing and developed world (Santanu et al., 2012). Alcohol abuse can cause many diseases, including; cancer of the mouth, esophagus


Introduction
Tobacco and alcohol consumption play an important role in mortality and morbidity in various countries (Ahmed, et al., 2003;Lopez, et al., 2006;Begg et al., 2007;Ahmed, et al., 2010).Globally, tobacco consumption is the main cause of death, resulting in millions of deaths every year, more than HIV/AIDS, tuberculosis and malaria (WHO, 2008).Cigarette and Shisha smoking are important public health concern worldwide causing many diseases including; pulmonary disease, various cancers including those of the respiratory, digestive, and genitourinary systems, some leukemias and premature death (Office of the Surgeon General, 2004; Babiker et al., 2012).
Socioeconomic factors have strong impact on tobacco use worldwide (Bobak et al., 2000;Ahmed and Mahgoob, 2007).A strong inverse relationship between socioeconomic status and smoking exists, particularly in the develop world.Such relationship indicating that the poorest and least educated populations are more likely to smoke than the other counterparts (Laaksonen et al., 2005;Wipfli and Samet, 2009).
Alcohol consumption is associated with high mortality and morbidity in both developing and developed world (Santanu et al., 2012).Alcohol abuse can cause many diseases, including; cancer of the mouth, esophagus and larynx, liver cirrhosis, and pancreatitis, social consequences, such as family and workplace-related problems, road-traffic accidents, and interpersonal violence (WHO, 2002;Ahmed et al., 2013).As a result, alcohol use has been getting more public and research attention in recent years (Ahmed, 2013).
It is evident that the prevalence of tobacco consumption is rising in the Hail region.Female participation in tobacco and alcohol related studies in the Hail -Saudi Arabia represent a major obstacle since it is considered as social stigma due to complete prohibition by law (Ginawi, 2013).Despite good knowledge about the hazards of tobacco consumption, about 25% of the medical students were found to smoke (Al-haqwi et al., 2010).Despite the strong religious belief in Saudi Arabia against the use of alcohol and other addictive substances, a great majority of the medical students in Saudi Arabia, perceived that alcohol abuse is a common problem in the community (Al-haqwi, et al., 2010).Cigarette smoking is an important public health problem in Saudi Arabia.A more intense and comprehensive tobacco control effort is needed (Jarallah, et al., 1999).Cigarette smoking is prevalent among Saudi adults in Riyadh, particularly males, most of whom begin to smoke rather early in life and continue for many years (Saeed et al., 1996).
The current study was carried out with the aims to assess the frequencies of tobacco smoking and alcohol consumptions among different population settings in Northern Region of Saudi Arabia (Hail Region) to provide data, which help in future planning to eliminate these habits, particularly among young generations.

Materials and Methods
This is a community-based, cross-sectional study, which was conducted in North Saudi Arabia, during the period from October 2014 to January 2015.Data was collected as a part of cancer awareness campaign that covered an area inhibited with approximately 140,000 individuals.People were gathered in certain public entities (University colleges, Health centers, Community gatherings), then were requested to fill a questionnaire about tobacco and alcohol habits and other information regarding their attitudes towards these issues.
Buccal cells were obtained by scraping buccal mucosa for subsequence molecular analysis of P53 gene.P53 gene mutation was examined using Polymerase Chain Reaction (PCR).
Statistical analysis: Statistical analysis was performed by proportion.The SPSS software (version 16) was used for statistical analysis.
Ethical consent: A written informed consent was acquired from each respondent, ensuring strict anonymity.
Table 4 summarizes tobacco and alcohol habits by occupation.Since, students representing the great majority of the study subjects, elevated number of smokers was found amongst students group representing 235 followed by employee, others, teachers and medical constituting 214, 102, 37, and 30 respectively.Regarding alcoholic, similarly most users were found amongst students constituting 14 followed by employees and others, representing 13 and 8, in this order.However, when calculating the percentages within each entire occupational group, the percentages greatly differ from the proportions of numbers of users within the study population as indicated in Figure 3.

Discussion
The tobacco used in a particular product has an important effect on its chemical composition, and differs with tobacco species, growing, curing, processing and storage.Therefore, it differs in its broad health consequences including cancer.Moreover, the epidemiology of tobacco use vary from country to another and from community to another.Evaluating cancer awareness is an important step toward cancer control; as it gives the policy makers and health care providers an opportunity to recognize the influence of the interventions and do what is possible to get the anticipated outcomes.Therefore, the aim of this study was to carry out an epidemiological survey on tobacco and alcohol use in Northern Kingdom of Saudi Arabia (Hail Region), which is an area inhibited by approximately 500,000 persons.
Although, there a number of studies performed in this context, bust mostly included younger population, which is differ in this study.Of these studies, the study by    (Alshammari, et al.,2015).The association between tobacco use and cancer was well well-known.Studies published since the 1986 IARC Monograph on "Tobacco smoking" The relationship between cigarette smoking and cancer of the lung, nasal cavities and paranasal sinuses, nasopharynx, stomach, liver, kidney etc., was strongly evidenced (Sasco, et al. 2004).
Nevertheless, this study investigated the presence of P53 mutation among tobacco users.The relationship between p53 gene mutations, tobacco smoke, and alcohol consumption was established in many cancers (Gerd, et al., 2002).Although, in this study there was no P53 mutation detected, there were many studies in this context.In a study examined p53 gene mutations in exons 5 through 8 by PCR-single-strand conformation polymorphism analysis in a cohort of 84 patients with laryngeal squamous cell carcinoma, they found p53 gene mutations were detected in 24 (28.6%)cases (95% confidence interval, 19.3%-39.5%),and the GC to TA transversion (33%) was the most common type of mutation (95% confidence interval, 15.6%-55.3%).Most mutations mapped to the p53 DNA-binding domain, which is necessary for the physiological activity of p53 as a tumor suppressor.A statistically significant association was found between p53 mutations and exposure to tobacco smoke (P=.001), which was the only variable significantly associated with p53 mutations in a multivariate model.The association with alcohol consumption was only at a borderline level of significance (P=0.065)(Ronchetti, et al., 2004).
Nevertheless, it was observed that in the present study many study subjects were living with smokers and this explore the burden of tobacco use which in most instances is considered as social stigma, particularly among females.Moreover, the stigma is at its highest levels among females which explain the small number of females' users in this study.
However, statistical figures regarding variables such as age, education and occupation have varies conditions and they were included to show the level of differences in order to stimulate further research in such context.
A limitation of the present study include the fact that data were collected via self-report.The of quantitative measures of tobacco exposure such as exhaled carbon monoxide, urine or saliva nicotine would allow a more conclusively categorized tobacco exposure in such population.Ways should be searched to come over stigma, which give rise inaccurate data.
In conclusion: Tobacco use and exposure are prevalent in Northern KSA, which require intervensions and controls.Alcohol use has low prevalence.No relationship was identified between P53 mutation and Tobacco use.

Figure
Figure 2. Description of the Study Subjects by Education and Habits within Entire Education Level

Table 3
Summarizes, tobacco and alcohol habits among different education levels.Individuals at university levels represent the highest proportions in smoking, and alcohol