Introduction of apoptosis of human hepatoma cells HEPG-2 with liposome-mediated c-myc antisense phosphorothioate oligodeoxynucleotide and 5-FU

Abstract

Objective To study the inhibition effect of c-myc ASODN (antisense oligodeoxynucleotide) and 5-FU (5-fluorouracil) on the expression of c-myc gene and the proliferation of human hepatoma cells HEPG-2. Methods After treated by liposome mediated c-myc antisense phosphorothioate oligodeoxynucleotide (APSODN) and 5-FU, the growth inhibition rate was detected by MTT assay, the expression of c-myc mRNA was detected by RT-PCR and immunohistocehemical methods HEPG-2cells. The cell cycle was analyzed by flow cytometric analysis. The morphological changes were observed by fluorescence staining and cellular genome electrophoresis. Results After sealing c-myc gene with ASODN,the growth of cells was repressed and the effect was time-dependent and dose-dependent ( P = 0. 02 ). The ability of proliferation decreased, the expression of c-myc gene was inhibited on transcription and translation levels; 5-FU can induce apoptosis of hepatoma cells HEPG-2 dramatically with the dose of 10 μ mol/L, when treated by both c-myc ASODN and 5-FU, HEPG-2 cells was induced apoptosis in a cooperative style ( P =0. 01 ). Conclusions The liposome mediated c-myc (APSODN) and 5-FU can inhibit the proliferation of HEPG-2 cells by inhibiting the expression of c-myc gene and can induce apoptosis of hepatoma cells HEPG-2 in a cooperative style. c-myc ( APSODN ) can increase the sensitivity of 5-FU to hepatoma cells and decrease the effective concentration of 5-FU. Key words: Carcinoma, hepatocellular; Apoptosis; Gene,c-myc