Introduction of apoptosis of human hepatoma cells HEPG-2 with liposome-mediated c-myc antisense phosphorothioate oligodeoxynucleotide and 5-FU
Tiankang Guo, Hui Cai, Yanqing Sun, Zhenjun Li, Wei Xiao Li
Abstract
Tiankang Guo, Hui Cai, Yanqing Sun, Zhenjun Li, Wei Xiao Li
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
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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
Key concepts: Apoptosis, Molecular biology, Liposome, Gene expression, Cell growth, Messenger RNA, Growth inhibition, Gene