L Tumanova, V Mitin, N Godoroja, N Botnariuc
breast cancer, fibrosing adenomatosis, gardnerella vaginalis, pcr
L Tumanova, V Mitin, N Godoroja, N Botnariuc. Gardnerella vaginalis and breast cancer. The Internet Journal of Oncology. 2008 Volume 6 Number 2.
Bacterial infections traditionally have not been considered major causes of cancer, however, a number of studies [1, 2, 3] show that some bacteria might be associated with cancer. We suggested that bacterial infection could be one of the reasons for breast cancer incidence. Preliminary PCR-analysis of tissue, blood and saliva samples of 10 breast cancer patients for such common pathogens as
Materials and methods
Breast disease (BD) group consisted of 41 patients of the Institute of Oncology of Moldova, who were recommended tumor resection, according to the primary diagnosis. There were no further restrictions when organizing this group. After the confirmation of the diagnosis, this group was divided into two subgroups: cancer group – 31 patients and fibrosing adenomatosis (FAM) group – 10 patients. The cancer group itself was divided into four subgroups, according to cancer stage (4 patients with T1 cancer stage, 19 patients with T2 stage, 4 patients with T3 stage, and 4 patients with T4 stage). The cancer stage was determined according to the International Classification of Diseases for Oncology, WHO Geneva (ICD-0-C50).
All patients were informed about the conducted research, and gave a written consent for it, in accordance with the legislation of the Republic of Moldova (The Law of Patient’s Rights in Biomedical Research).
Control group consisted of 30 healthy individuals (13 individuals from Institute of Genetics and Plant Physiology and 17 from Institute of Oncology). Healthy status for the control group was determined by questionnaire.
Uro group was formed based on retrospective studies of 40 patients (21 women, 19 men), who sought medical help for urogenital diseases . The probabilities of
Blood and saliva collection from BD group patients was performed in the Institute of Oncology of Moldova 1 – 2 hours before tumor resection, and tissue samples – after tumor resection. In case of the control group, only saliva was collected for the study. All samples were stored and transported at +4°C. Time between sample collection and DNA isolation did not exceed 24 hours.
DNA isolation and PCR-analysis.
DNA from blood and saliva was isolated using lysis buffer containing 5M guanidinium thiocyanat, 50mM Tris (pH 7.5), 10mM EDTA, 0.5% Triton X-100, then extracted using equal volume of phenol:chlorophorm (vol/vol) and then chlorophorm. DNA was precipitated by 2.5 volumes of ice-cold 96% ethanol in the presence of 0.3M ammonium acetate. Tumor was grinded in liquid nitrogen with addition of lysis buffer, and then extracted as above.
To improve the sensitivity of
As a positive control DNA template of
Odds ratio (OR) and relative risk (RR) with 95 % CI (Confidence Interval) was carried out using medcalc software (http://www.medcalc.be/calc/odds_ratio.php). We calculated the Yates-corrected chi-square (χ2) and 95% CI, according to Glantz , P
Fig.1A shows an example of
Table1 shows the results of
A – frequency of bacterial detection in breast disease group for each of the analyzed samples (b, s, t,), as well as for the combinations (bs, bst); B - distribution of positive cases (bs) in different patient groups and control group .
The frequency of bacterial detection either in blood (b) – 43%, or in saliva (s) – 46% is practically similar, and increases substantially (1.6 times) when both these data (bs) are considered. The probability of bacterial detection further increases, if
Data presented in the given research demonstrate a high frequency of
Of importance is a rather high frequency of
Application of polychemotherapy for the patients of T2-T4 groups could lead to decrease of concentration or disappearance of
In this research, the results of