engleski

Testing of Atrazine Cytotoxicity on Chinese Hamster Ovary (CHO-K1) Cell Line

Atrazine, as s-triazine herbicide, selectively controls broadleaf weeds and certain grass weeds in corn and sorghum fields. An estimated 34, 5 million kg of atrazine are applied to cropland each year in the United States, and it is also one of the most used herbicides in Europe. The US EPA does not consider likely the atrazine to be a human carcinogen. However, its effects on other physiological processes point to atrazine as an endocrine disruptor. In order to determine possible toxic effect at ovarian cellular level, in this study Chinese Hamster Ovary (CHO-K1 ; CCL-61) cell line was used. Cells grew in monolayer at 37 &ordm ; C in the atmosphere of 95% air and 5% CO_2, in the medium Dulbecco's MEM with 5% newborn calf serum. The biomass was produced in T-bottles. Cells were separated in the early logarithmic phase of growth and seeded on multiwell plates in concentration of 1, 5 x 10^4 cells/mL of cultivating medium. Cytotoxic effect of atrazine at the concentration range of 10 – 160 micro g/mL was determined by colorimetric methods after 24, 48 and 72 hours. IC_50 value was determined from the slope of % inhibition vs. log dose values. Cell viability and the number of cells were measured by: Trypan Blue exclusion method (IC_50 95, 3 &plusmn ; 9, 25 micro g/mL) ; Kenacid Blue R binding method change in total cell protein (IC_50 173, 85 &plusmn ; 13, 35 micro g/mL) ; lyzosomal activity by Neutral Red method (IC_50 196, 24 &plusmn ; 9, 8 micro g/mL) ; MTT assay, the ability of viable cells to convert a soluble tetrazolium salt into insoluble formazan precipitate (IC50 157, 17 &plusmn ; 14, 14 micro g/mL). Application of in vitro assays in evaluation of the toxicity should be an important contribution in elucidating cellular and molecular mechanisms of atrazine.

atrazine; CHO-K1 cell line; cytotoxicity

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