Changes of hepatocellular carcinoma cell structures during apoptosis induced by Survivin antisense oligonucleotide / 中国普通外科杂志

ABSTRACT

ObjectivesTo investigate the changes of hepatocellular carcinoma cell line SMMC-7721 in cell ultrastructure and skeleton during apoptosis induced by Survivin ASODN.MethodsSurvivin ASODN was transfected into SMMC-7721 cells mediated by DOTAP liposomal reagent. The changes of Survivin protein and mRNA expression after transfection were assessed by Western blot and in situ hybridization. The apoptotic rate was detected by flow cytometer. Cell ultrastructure and skeleton were observed by electron microscope and confocal microscope. Caspase-3 was assessed by kinase activity assess method. Results The expression of Survivin protein and mRNA decreased from 69.59 and 75.6 to 10.71 and 22.9 respectively. The cell ultrastructure showed apoptotic changes. Cell skeleton microfilament system changed after Survivin ASODN transfection. The average intensity of microfilament system fluorescent decreased from 190 to 64. The activity of Caspase-3 increased from 0.015 3 to 0.099 2.ConclusionsTransfection of ASODN targeted to Survivin mRNA by DOTAP liposomal transfection reagent down-regulated the expression of Survivin protein and mRNA in SMMC 7721 cell line and activate Caspase-3, which in turns induced cell apoptosis.