engleski

Different Apoptosis Ratio and Gene Expression in Two Human Cell Lines after Sevoflurane Anaesthesia

Background: Sevoflurane is an inhaled anesthetic often used alone or as a part of balanced anesthesia. The aim of this study was to determine the consequence of single exposure of carcinoma (Caco-2 and HEp2) cells to clinically utilized gas mixture containing sevoflurane 3 vol% on apoptosis induction, propapototic genes expression, and activation of sphingomyelinase apoptotic pathway. Methods: Apoptosis was determined by flow citometry. P53, caspase 3, and CYP2E1 gene expression was determined using RT-PCR. Activities of acid (aSMase) and neutral (nSMase) sphingomyelinases were measured using methyl-14C sphingomyeline, and for de novo ceramide and lipid synthesis [3H] palmitic acid was used. All results were compared with controls and analyzed by Mann-Whitney and Kruskal-Wallis tests. Results: In treated Caco-2 cells increased amount of apoptotic cells (16.9% ; p=0.04) 24 h after exposition. Expression of p53 and caspase-3 genes rose in Caco-2, and decreased in HEp-2 cells. The CYP2E1 gene expression was observed in the Caco-2 cells only. In control cells catalytic activity of aSMase was higher than nSMase activity for 2.3-fold. Decreased aSMase and nSMase activities were observed in Caco-2 cells 24 h after exposition. ASMase activity was halved (54.2% ; p=0.06) in HEp-2 cells 24 h after anaesthesia. De novo ceramide synthesis correlated SMase activities in Caco-2 cells. Conclusion: Sevoflurane anaesthesia induces late apoptosis in investigated colon and laryngeal carcinoma cells. Although obtained results lead us to conclude that in the Caco-2 cells anaesthetic gas mixture containing sevoflurane induces p53-dependent apoptosis, the mechanism of apoptosis induction is still unclear and remains to be elucidated.

Sevoflurane; human tumour cells; apoptosis; p53; CYP2E1; sphingomyelinase activity

nije evidentirano