Different apoptosis ratios and gene expressions in two human cell lines after sevoflurane anaesthesia.
BACKGROUND
The aim of this study was to determine the effect of a single exposure of carcinoma cells (Caco-2 and HEp-2) to an anaesthetic gas mixture containing sevoflurane 3%, applied for a period of either 1 or 2 h, on the induction of apoptosis, propapototic gene expression and sphingomyelinase activity.


METHODS
Apoptosis was determined by flow cytometry. p53, caspase 3 and CYP2E1 gene expression was determined using reverse transcriptase polymerase chain reaction. Activities of acid (aSMase) and neutral sphingomyelinases (nSMase) were measured using methyl-(14)C sphingomyeline, and for de novo ceramide and lipid synthesis [(3)H] palmitic acid was used. All results were compared with controls and analysed by Mann-Whitney and Kruskal-Wallis tests.


RESULTS
In the treated Caco-2 cells, the apoptotic ratio increased 24 h after anaesthesia (16.9%; P=0.04). The expression of both p53 and caspase-3 genes increased in Caco-2 and decreased in HEp-2 cells. The CYP2E1 gene expression was observed only in the Caco-2 cells. In control cells, the catalytic activity of aSMase was 2.3 times higher than that of nSMase activity. 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 with SMase activity in Caco-2 cells.


CONCLUSIONS
Sevoflurane anaesthesia induces late apoptosis in the colonic and laryngeal cancer cells investigated. Although the results obtained may indicate that an anaesthetic gas mixture containing sevoflurane induces p53-dependent apoptosis in the Caco-2 cells, the mechanism of apoptosis induction is unclear and remains to be elucidated.
Kvolik
S
S
Department of Anaesthesiology and ICU, University Hospital Osijek, Osijek, Croatia. slavica.kvolik@os.t-com.hr
Dobrosevic
B
B
Marczi
S
S
Prlic
L
L
Glavas-Obrovac
L
L
D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics*; D045744:Cell Line, Tumor; D002518:Ceramides / Q000378:metabolism; D019392:Cytochrome P-450 CYP2E1 / Q000096:biosynthesis / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects*; D006801:Humans; D050356:Lipid Metabolism / Q000187:drug effects; D008738:Methyl Ethers / Q000494:pharmacology*; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077149:Sevoflurane; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism
D018685:Anesthetics, Inhalation; D002518:Ceramides; D008738:Methyl Ethers; D000077149:Sevoflurane; D012313:RNA; D019392:Cytochrome P-450 CYP2E1; D013108:Sphingomyelin Phosphodiesterase