Hepatoma tissue culture (HTC) cells as a model for investigating the effects of low concentrations of herbicide on cell structure and function
Abstract
Previous studies on mice fed genetically modified (GM) soybean demonstrated modifications of the mitochondrial functions and of the transcription/splicing pathways in hepatocytes. The cause(s) of these alterations could not be conclusively established but, since the GM soybean used is tolerant to glyphosate and was treated with the glyphosate-containing herbicide Roundup™, the possibility exists that the effects observed may be due to herbicide residues. In order to verify this hypothesis, we treated HTC cells with 1–10 mM Roundup and analysed cellular features by flow cytometry, fluorescence and electron microscopy.
Under these experimental conditions, the death rate and the general morphology of HTC cells were not affected, as well as most of the cytoplasmic organelles. However, in HTC-treated cells, lysosome density increased and mitochondrial membranes modified indicating a decline in the respiratory activity. Moreover, nuclei underwent morpho-functional modifications suggestive of a decreased transcriptional/splicing activity.
Although we cannot exclude that other factors than the presence of the herbicide residues could be responsible for the cellular modifications described in GM-fed mice, the concordance of the effects induced by low concentrations of Roundup on HTC cells suggests that the presence of Roundup residues could be one of the factors interfering with multiple metabolic pathways.
Keywords: Cell nucleus; Cytometry; Electron microscopy; Fluorescence microscopy; Herbicide; HTC cells; Lysosomes; Mitochondria
Abbreviations: DFC, dense fibrillar component; EDTA, ethylene diamino tetraacetic acid; FC, fibrillar centre; GC, granular component; GM, genetically modified; HTC, hepatoma tissue culture; NGS, normal goat serum; PBS, phosphate-buffered saline; PG, perichromatin granule