ABSTRACT
AIM:To observe the effect of antisense locked nucleic acid (anti-LNA) blocking the translation initiation region of c-myc exon 2 on the viability and apoptosis of hepatocellular carcinoma cells.METHODS:The antiLNA that was complementary to the translation initiation region of c-myc exon 2 was designed,synthesized,and introduced into the HepG2 cells by cationic liposome-mediated transfection.The mRNA and protein levels of c-Myc in the cells were determined by RT-PCR and Western blot.The change of cell apoptosis was analyzed by flow cytometry,and the toxicity of anti-LNA to the cells was detected by MTT assay.RESULTS:Five days after transfection,the mRNA level of c-Myc in anti-LNA group was 0.335 ±0.016,and the protein level was 0.448 ± 0.037,significantly lower than those in control group (both P ＜ 0.05).The ratio of apoptotic cells in anti-LNA group was 32％ ±-6％,which was higher than that in control group (P ＜ 0.05).CONCLUSION:Antisense locked nucleic acid targeting at the translation initiation region of cmyc exon 2 shows strong inhibitory effects on the apoptosis of hepatocellular carcinoma cells.
ABSTRACT
AIM:To observe the effect of antisense locked nucleic acid (anti-LNA) blocking the translation initiation region of c-myc exon 2 on the viability and apoptosis of hepatocellular carcinoma cells.METHODS:The antiLNA that was complementary to the translation initiation region of c-myc exon 2 was designed,synthesized,and introduced into the HepG2 cells by cationic liposome-mediated transfection.The mRNA and protein levels of c-Myc in the cells were determined by RT-PCR and Western blot.The change of cell apoptosis was analyzed by flow cytometry,and the toxicity of anti-LNA to the cells was detected by MTT assay.RESULTS:Five days after transfection,the mRNA level of c-Myc in anti-LNA group was 0.335 ±0.016,and the protein level was 0.448 ± 0.037,significantly lower than those in control group (both P ＜ 0.05).The ratio of apoptotic cells in anti-LNA group was 32％ ±-6％,which was higher than that in control group (P ＜ 0.05).CONCLUSION:Antisense locked nucleic acid targeting at the translation initiation region of cmyc exon 2 shows strong inhibitory effects on the apoptosis of hepatocellular carcinoma cells.