ABSTRACT
Objective Proliferation and apoptosis play a major role in the development of tumor cells,and the intranuclear transcriptional factor c-fos is significantly up-regulated in the primary hepatocellular carcinoma and involved in early carcinogenesis.The purpose of the present study is to investigate the apoptotic effect of c-fos antisense oligodeoxynucleotide(ASO) on human hepatocellular carcinoma HepG2 cells and the participation of Caspase3 in this process.Methods Cell culture,Hoechst 33258 staining,real-time PCR and Western blotting were used in present study.Cultured HepG2 cells were divided into 3 groups: 1) control group: cultured with 10?l saline;2) sense oligodeoxynucleotide(SO,used as a negative control) treated group: co-cultured with 10?l SO(5?g/?l);3) ASO treated group: co-cultured with ASO 10?l(5?g/?l).The subsequent experiments were performed 1h after cultivation for each group.Hoechst 33258 staining was performed to detect the apoptosis by observing the staining of nuclear chromatin.Real-time PCR and Western blotting were used respectively to detect the expression of Caspase 3 at mRNA and protein levels after different treatments.Results Hoechst 33258 staining revealed that the nuclei of HepG2 cells showed diffuse and adqulis fluorescence in control and SO-treated groups,while dense and dark fluorescence was observed in ASO-treated group,which indicated that c-fos ASO had significantly induced apoptosis of HepG2 cells.The expression of Caspase 3 in ASO group was enhanced both at mRNA and protein levels compared to that in control groups.Conclusions C-fos ASO significantly induces apoptosis in human hepatocellular carcinoma HepG2 cells,as shown by Hoechst 33258 staining and higher expression of Caspase 3 mRNA and protein.Moreover,Caspase 3 activation is involved and probably plays an important role in c-fos ASO-induced apoptosis of HepG2 cells.