@2024 Afarand., IRAN

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ISSN: 2251-8215    Sarem Journal of Reproductive Medicine 2017;1(3):89-95

Background
Breast cancer is the most common malignancy in women [1]. ... [2-4].In developing countries, the ‎incidence of breast cancer is also increasing [5].‎
Previous Studies
‎.. [6]. Laboratory tests are one of the most important and most effective early diagnostic tools that are ‎based on the evaluation of markers in available body fluids such as peripheral blood. One of the ‎indicated markers is the radiation sensitivity of the lymphocyte.‎ Chromosomes tend to be sensitive to radiation in most cancers, such as breast cancer, a percentage of ‎colorectal cancer, or cancers of the head and neck. About 5 to 14% of the natural population is also ‎allergic to radiation. Therefore, it has been suggested that chromosomal sensitivity to radiation be ‎used as an indicator of the potential for cancer. Differences in the ability to repair DNA may cause a ‎change in radiation sensitivity among normal individuals [5].‎
Aim(s)
The aim of this study was to evaluate the sensitivity of radiation in non-inherited breast cancer in ‎Iranian women.‎
Research type
This study is an observational-analytical study carried out using a case-control method.‎
Research society, place and time
Individuals with non-inherited breast cancer were examined at Mehrad Hospital in Tehran compared ‎with healthy people.‎
Sampling method and number
A total of 32 individuals with non-inherited breast cancer were studied. These patients did not receive ‎any treatment at the start of the study. Thirty healthy subjects were selected as control group, who had ‎no history of cancer in their first and second degree relatives. Since access to healthy volunteers who ‎had all criteria for entering the study, along with the criteria for matching on the basis of the variables, ‎were limited, in some variables, such as age at the first pregnancy, menarche age and age of ‎menopause, the two groups were identical and in Others, such as the age and the number of ‎pregnancies, this consistency was not achieved that with the help of statistical evaluations, the effect of ‎these variables was checked so that, if there was an effect on the outcome, it could be counterbalanced.‎
Used devices&materials
Before sampling, consent was taken from all the participants. 5 cc of Heparin blood was taken from all ‎the participants. Samples were left at 4 ° C for 24 hours and then cultured. In the patient group, all ‎clinical parameters such as tumor size, lymph node involvement status, estrogen receptor and ‎progesterone receptor status, status of expression of p53, HER-2 and grade stage genes were studied ‎and recorded through patient files.‎ The RPMI 1640 culture media which contained bicarbonate sodium and L-glutamine (Eurocolone Lot ‎No: ECM 0495D), penicillin-streptomycin (Eurocolone Cat No: ECB3001D), Eurocolone Lot No: ‎ECS0170D and phytohemagglutinin (GIBCO Cat No: 10576- 015) were used. According to the ‎chromosomal decomposition program using G2, irradiation was performed 72 hours after cell culture. ‎The incident beam was gamma ray with a dose of 0.4 Gy.‎ Removal of cells: Half an hour after radiation by adding calcium, the cells stopped in the metaphase ‎step [7-15]. The second step was to adjoin them with a hypothalamic solution (Hypotone). Due to this ‎proximity, the cells were bulky and the chromosomes got enough space to disperse within the ‎lymphocytes. The third stage is the stabilization of the cells. Finally, samples were prepared and ‎examined after staining. From each sample, three cultures were prepared, two of which were exposed ‎to radiation, and third culture as control culture in all conditions was similar to the other two cultures ‎except irradiation. From the irradiation cultures, a total of 50 cells were examined and from control ‎culture, also, 50 cells were removed for chromosomal counts and evaluation.‎ In the microscopic analysis of metaphyseal lymphocytes, the percentage of abnormal cells was used to ‎determine the sensitivity of the radiation. Maladaptive cells are counted based on the existence of ‎failures that cause abnormalities, chromatid and chromosome clefts (when lack of continuity in the ‎chromatid building is wider than the width of a chromatid), rearrangements (such as chromosomes ‎ring and dicentric chromosome) and the radial forms (3 radial and four radial forms) cells. When ‎discontinuity in the structure of the chromatid is less than one chromatid, chromatid and ‎chromosomal clefts are not considered as chromosomal abnormality.‎ Data were analyzed by SPSS 19 software. Parametric tests (T-test and Paired t-test) and nonparametric ‎tests (Chi-square and Fisher exact tests) were used to assess the significant difference between mean ‎sizes of variables. The evaluation of the radiation sensitivity test was performed based on the ‎characteristics of the system function or the ROC Curves 1 and performing the C-statistics. In this ‎study, in addition to calculate the borderline for each test index based on the Youden index, the ‎sensitivity and specificity of the test were determined at the critical point and the odds ratio was ‎estimated at 95% confidence interval.‎
Findings by Text
The mean age in the affected population was 53.25 years with a range of 32-84 years and in healthy ‎subjects it was 31.33 years with a range of 18-49 years (Table 1).‎ Considering the significant difference in the age and number of pregnancy variables between the two ‎groups of patients and control, it is likely that these two variables can be considered as confounding ‎ones and this can cause the results to be skeptical, that in this respect, the necessary studies were ‎done. Since for a variable to be confounding, the existence of a relationship with both the independent ‎variable and the dependent variable or the same outcome is certain, it should be said that the two ‎variables of age and the number of pregnancies, at least with the condition of exposure to radiation ‎sensitivity, had no definable and definite association. However, statistical analysis took place in this ‎regard. Pearson and Spearman correlation test showed a lack of correlation with the results of the tests ‎and, therefore, there was no ambiguity in term of their likelihood of confusion. ‎ The distribution of breast cancer patients according to the tumor stage (according to the three-letter ‎TNM system) showed that 7 patients (23.3%) had stage I, 4 cases (13.3%), had stage Iia; 16 cases ‎‎(53.3% %) had Stage IIb, one case (3.3%) had stage IIIa and two cases (6.7%) also had stage IIIb. In ‎‎62.5%, the tumor had a moderate distinction or a score of 2. In 33.3% of the cases, the tumor had a ‎high distinction or score of 1, and only in 3.1 % of the patients, tumor had a score of 3. The presence or absence of estrogen, progesterone, p53 and HER-2 receptor was a very important part ‎of the study (Table 2).‎ The percentage of abnormal cells in both patient and control groups was significantly different in each ‎case before and after irradiation (Chart. 1, p = 0.00001).‎ Comparing this index between patient groups and the control group, the presence of abnormal cells ‎prior to irradiation showed no significant difference (p> 0.05), but after irradiation, and also the ‎difference in pre and post changes in the percentage of abnormal cells, the difference was significant ‎‎(p<0.05).‎ Statistical analysis and calculation of C index for percentage of abnormal cells in G2 stage to γ radiation ‎showed that this index can be a significant index with acceptable level of AUC (0.725) (p <0.05) that ‎can be helpful in terms of clinical and decision-making with the odd ratio (Chart 2).‎ The odds ratio at critical point of 61% was 3.818 (CI95% = 1.323-10.942), so that in those who had an ‎increase in the percentage of abnormal cells after irradiation than before irradiation it was equal to or ‎greater than 61%. The probability of being affected was more than 4 times (3.818) more than other ‎people. At the critical point of 61%, the sensitivity of this index was 65.6% and the specificity was ‎‎66.6%, which could be acceptable for a test. This test can accurately identify approximately 65.6% of ‎the patients and 66.6% of normal people can be identified as healthy with test. ‎ Irradiation caused various clefts and abnormalities in chromosomes (Figures 1 and 2).‎
Main comparison to the similar studies
‎... [16-19].In various studies, the sensitivity of radiation in patients with breast cancer has been ‎investigated [14, 15, 20-22]. In some studies, it has shown that about 40% of breast cancer patients ‎have high sensitivity to radiation [8, 12, 13, 23-26]. The sensitivity of the patients to radiation in this ‎study was in line with expectations and in line with other studies. Recently, several studies have tried to address the issue of chromosomal sensitivity at the cytogenetic ‎molecular level and the correlation of the outcome (breast cancer disease) with various characteristics ‎such as genetic variations and even polymorphisms [27-29].‎ ‎... [30, 31]. Research results based on new technology in Ukraine are consistent with our research ‎findings regarding the feasibility and applicability of radiation sensitivity testing in the G2 phase of the ‎cell cycle (with X-ray radiation, however) [32]. ... [33-35].‎
Suggestions
The use of the chromosomally sensitive radiation test can be more accurately assessed in cohort ‎studies in the future.
Limitations

Conclusions
Iranian women with non-inherited breast cancer have more susceptibility to gamma radiation than ‎healthy women, and show this sensitivity as chromosomal breaks and clefts. Therefore, ‎chromosomally sensitive radiation test, have the usability as an early detection of breast cancer ‎biomarker, or a possible indicator of the potentiality of being affected with this disease.
Acknowledgments
The Genetics Research Center of the University of Rehabilitation Sciences and Welfare is appreciated ‎for financial support. The staff of the Mehrad Hospital are appreciated for their sincere help in taking ‎samples from the patients in optimal condition. Staff members of the Radiation Therapy Center, are ‎thanked and appreciated who helped us to irradiate the samples, and all those who collaborated with ‎us in this project are appreciated as well.
Conflict of interest

Ethical Permissions

Funding Sources
This research was funded by the Genetics Research Center of the University of and Rehabilitation ‎Sciences and Welfare.‎

TABLES and CHARTS

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