engleski

Phytohaemagglutinin as a modulator of chromosomal aberration assay and micronucleus assay in ionizing radiation biodosimetry

For more than 30 years chromosomal aberration analysis and micronucleus assay have been considered to be important cytogenetical tool in biological effects evaluation of ionizing radiation. To make the radiation-caused DNA damage visible, cells have to be initiated to enter the G1 phase, to pass the S phase of the cycle.Therefore the cultivation of peripheral blood lymphocytes in the presence of mitogen activator is required. The mostly used is phytohaemagglutinin. We intended to find out whether the point when the phytohaemagglutinin is added after the culture initiation has any influence on chromosomal aberration and micronuclei formation. Human peripheral blood lymphocytes were isolated. To initiate DNA damage lymphocytes were irradiated by 2 Gy using a gamma-ray 60 Co source. The cultures for chromosomal aberration analysis and micronucleus assay were started and phytohaemagglutinin was added 0, 1, 2 or 4 hours after irradiation. The number of dicentric chromosomes and acentric fragments were significantly increased in lymphocytes stimulated by phytohaemagglutinin immmediately after irradiation compared to the cultures where activator after 1, 2 and 4 hours was added. The number of aberrations between activator after 1, 2 and 4 hours after irradiation did not differ significantly. Micronucleus assay did not show any significant differences in number of micronuclei regardless of the time when the mitogen activator was added. We could say that in order to maximize the sensitivity of chromosomal aberration analysis phytohaemagglutinin has to be added immediately after the irradiation.

Ionizing radiation; Phytohaemagglutinin; Chromosome aberration; Micronucleus assay

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