Číslo abstraktu: p008
Aims: the aim of this research was to determine metallothionein levels in human fibroblast cell cultures (derived from an individual with cancer) exposed to cadmium.
Materials and methods: BR-175 (tumour) and BR-142 (control) human fibroblast cell cultures were exposed to CdNO3 in concentrations 0, 1, 5 and 10 uM. In intervals of 0, 1, 3, 6, 12 and 24 hours the cells were harvested. MT level was determined by using Western blotting, Dot-Immunobinding Assay (DIA) and Differential-Pulse Voltammetry-Brdicka reaction (DPV).
Results: MT levels determined in studied cell lines were within the range from 180 to 48.900 ug/mg of the soluble proteins. We observed a different behaviour of tumour cells in response to heavy metal exposure compared to controls. In control cells (0 uM Cd2+) MT levels were 2-3 mg/g of soluble proteins and did not change over time compared to treated cells, in which we observed a marked enhancement of MT level even after one hour long treatment. Compared to controls (BR 142) the MT level was three-times greater in tumour cells (BR 175) at the beginning of the experiment. The increase in MT content in BR 142 cells exhibited a linear trend over time, while for BR 175 cells MT induction was faster, but with prolonged exposure, MT content in tumour cell was below 75 % compared to control cells in relation to Cd concentration.
Conclusions: The increase in MT synthesis and different kinetics of MT induction after cadmium exposure in tumour cells in comparison to controls, found by analytical methods used, indicates a possible role of MT in carcinogenesis and metastasis
Acknowledgement: This project was supported from grants Liga proti rakovine and KAN208130801.
Datum přednesení příspěvku: 24. 4. 2009