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Sevoflurane induces apoptosis and modulates expression of proapoptotic genes in human colon carcinoma cells

Our study was designed to determine the consequence of single exposure of carcinoma cells to sevoflurane, simulating thus the situation occurring in the clinical setting. Colon carcinoma cells (CaCo2) were exposed to anesthetic gas mixture consists of O2:N2O:CO2 (35:60:5 vol.%) and sevoflurane (3.0 vol.%) for 1 and 2 hours. Cytotoxicity was analyzed by MTT assay. Mode of cell death, caspase-3 enzyme activity, and expression of genes in control and treated cells were tested immediately after exposure to sevoflurane and 24 h later. Apoptosis was assessed by FACS analysis using annexin and propidium iodide staining. Caspase-3 enzyme activity was measured by EnzChek caspase-3 Assay Kit. Expression of genes (GADPH, p53, caspase-3, and CYP2E1) was examined by RT-PCR. Results of our study shown that sevoflurane significantly suppressed treated cells' growth. Percent of apoptotic and necrotic cells increased in comparison with control cells in a time-dependent manner. Alteration in caspase-3 enzyme activity was not observed. Compared to expression in control cells, caspase-3 gene expression changes (>1.5 fold) were found in treated cells at all time points. Expression of p53 gene immediately after treatment was unchanged and 24 h later significantly increased (for 78%). Expression of CYP2E1 gene also increased (for 73%) in the cell exposed to sevoflurane for 2 h, and 24 h later slow down to 20%. In conclusion, sevoflurane in anesthetic dose suppresses the colon carcinoma cells' growth and increases expression of the CYP2E1 gene, which could be involved in detoxification processes. Does sevoflurane induces apoptosis in CaCo2 cells by p53-dependent or p53-independent pathway remains to be identified.

inhaled anesthetics; apoptosis; expression of proapotptic genes

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