Abstract:

Objective To study the influence of inhibitor apoptosis protein (IAP) Apollon antisense oligodeoxynucleotide (ASO) on the proliferation and apoptosis on human liver cancer HepG₂ cells, and investigate the mechanisms. Methods Apollon ASO was incubated by liposome with human liver cancer HepG₂ cell for 48 h, and the proliferation inhibition was detected by WST-8. The expression of Apollon mRNA was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (PCR). The early apoptosis rates were determined by flow cytometry. The morphology of HepG₂ cell apoptosis was determined by Hochest 33258 staining. The cell apoptosis was determined by mitochondrial membrane potential. The activities of cysteine proteinases (Caspase-3 and Caspase-9) were determined by colorimetric method. The expression of cytochrome C was analyzed by Western blot. Results After being treated by Apollon ASO on HepG₂ cells for 48 h, it was found that Apollon ASO could all suppress the proliferation of HepG₂ cells, which was depended on the concentration. Apollon ASO could induce HepG₂ cell apoptosis. The different concentrations of Apollon ASO reduced significantly the expression level of Apollon mRNA. The activities of Caspase-3 and Caspase-9 and the protein expression levels of cytochrome C increased. Through observing by fluorescence microscopy assay, nucelus high fluorescence cells were found, and the apoptosis cytoryctes were also observed. Conclusions Apollon ASO can reduce the expression level of Apollon mRNA and inhibit the proliferation of liver cancer HepG₂ cells, and it can induce the apoptosis by the way of mitochondria mediation.

Key words: Antisense oligodeoxynucleotide, Apollon, Proliferation, Apoptosis, HepG₂ cell, Liver cancer