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Научни трудове на Съюза на учените в България-Пловдив, серия Б. Естествени и хуманитарни науки, т. XV, 2013 г. Научна сесия „Техника и технологии, естествени и хуманитарни науки", 25-26 X 2012 Scientific researches of the Union of Scientists in Bulgaria-Plovdiv, series B. Natural Sciences and the Humanities, Vol. XV, ISSN 1311-9192, Technics, Technologies, Natural Sciences and Humanities Session, 25-26 oktober 2012.
GENETIC DAMAGE OF MEDICAMENTS IN BUCALL CELLS AT PATIENT WITH LUNG CANCER
Shqiptar Demaqi1, Kemajl Kurteshi 2
faculty of Medicine, University of Prishtina, Kosovo 2 Department of Bilogy, Faculty of Natural Science, University of Prishtina, Kosovo
Abstract
Micronuclei frequency is a sensitive biomarker used to evaluate the genotoxicity induced by genotoxics.
The aim of the study is to estimate genotoxic effects of medicaments (which used for cure of tuberculosis) through micronucleus test in human buccal cells.
The chromosomal damage are analysed by micronucleus test in buccal epithelium cells collected from 25 patients from Pulmonary Clinics in Hospital of Prishtina (Kosovo).
The frequency of micronuclei in buccal cells was higher at patients treated with chimiotherapy compare with the healthy subjects, although the difference is significant.
The frequency of micronuclei in buccal epithelium of patients after exposure is higher than in control healthy subjects. This difference it's statistically significant (P > 0,001).
Keywords: Micronucleus, test, buccal cell, pulmonary clinic,
Introduction
Pulmonary cancer is among the ten types of malignant neoplasia of highest incidence worldwide and is particularly common in developing countries. Cigarette smoking is considered to be the most important risk factor for its development, particularly when in association with alcoholic beverages(Wunsch-Filho et al.2002, Rodriguez et al. 2004)
Similarl to other types of malignant neoplasia, pulmonary cancer results from alterations (point mutations and chromosomal abnormalities) in genes that control the cell cycle, and/or in genes that are involved in DNA repair.
In addition to the potential for metastasis, cancer is characterized by the loss of the ability of cells to evolve to death when genetic damage occurs (apoptosis),( Hanahan, 2000).
It has been shown that exposure of persons to many environmental and occupational pollutants can lead to increased level of MN in epithelial cells. On investigating the genotoxic effects of environmental factors on human nuclei, the micronucleus (MN) test has been employed on buccal epithelial cells (Sarto et al., 1987; Tolbert et al., 1992),
Material and methods
The study population consisted of 26 patient (13 males and 13 female) and control group of 13 healthy subjects. The youngest patient it's 17 year, and the oldest it's 71 year old. The patient
are subjected to chimiotherapy , with aim to cure the tuberculosis.
Buccal cells were sampled with toothbrush from the inside of the cheeks and placed in physiological solution(NaCl 0.9%). The cells were washed thrice in the buffer solution by centrifugation.
After centrifugation (10 min at 1000 rpm), the pellet was fixed in 3:1 methanol/acetic-acid, for 10 minutes.Cell suspension was dropped onto clean slides. Three slides were prepared for each subject and 1000 cells are scoring (at 100 x magnification), from each subject were examined. The cells were stained in 10 % Giemsa solution.
Results and disscusion
The results of the present work indicate that the consumption of medicament, damage the chromosomes, which damage we notice through the presence of MN in buccal cells.
The results we are presents in table 1. In table the results we separate according the gender and age.
The frequency of micronuclei in buccal cells after chemiotherapy at male and female is 7.19 MN/1000 bucal cell it was higher compared with MN at control group 1.28 MN/1000 bucal cell).
When we analysed the frequency of MN after separate the patient according the gender we found that the frequency of MN it was higher at male (7.33 MN/1000 cells) compared with female (6.23 MN/1000 cells), but not statistically significant.
Also frequnecy of micronuclei at male it was higher (7.33 MN/1000 cells)compared with control group (2.16 MN/cells) but not statistically significant.
Also and between female and control group are higher but not statistically significant(P=0.113)
The sick patients submitted to chimiotherapy were investigated cytogenetically after chimiotherapy exposure and 1000 buccal cells per sample were scored, wich is considered as an optimal number of cells to be scored for genotoxic effects estimation(Titenko-Holland et al.1994, Surrles et al.,1999).
A significant positive relationship with age has also been obtained by other authors investigating the spontaneous frequency of micronuclei in epithelial cells (Pastor et al.,2001 ) and peripheral lymphocytes(Thierens et al.,2000 , Fenech et al., 1986 )
Tab .1. Frequency of Micronuclei in patients after chemiotherapy, scored to 1000 buccal cells
Frequency of micronuclei(MN) at patient exposed to chemiotherapy Frequency of micronuclei at control group
Total number of micronuclei(female+male) 381 MN 41 MN
Mean value of MN 7.19 MN 1.28 MN
Total number of micronuclei at female 186 MN 40 MN
Mean value of MN at female 6.23 MN 1.33 MN
Total number of micronuclei at male 212 MN 49MN
Mean value of MN at male 7.33 MN 2.16 MN
Oldest group 7.6MN 4.9MN
Youngest group 5.13MN 2.63MN
Conclusion
According to this investigation , we can concluded :
-chemiotherapy induce increase the number of micronuclei in the buccal cells but not statistically significant compared with control group.
- Male has greater average number of MN (7.33 MN) compare with female(6.23) -Higher frequency of micronuclei at oldest group (7. 6 MN)compare with youngest group(5.13 MN), but not statistically significant.
-Age coorelates with micronucleus frequency -Sex not correlates with micronucleus frequency
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Рецензент: доц. д-р Ивета Коева
