论文部分内容阅读
目的探讨c-fos反义探针诱导人肝癌HepG2细胞凋亡以及caspase3在其中的作用。方法人肝癌HepG2细胞分为3组,①对照组:加生理盐水10μl;②c-fos正义寡核苷酸(SO)处理组:加入SO10μl(5μg/μl);③c-fos反义寡核苷酸(ASO)处理组:加入ASO10μl(5μg/μl)。各组细胞再培养1h后进行后续实验。采用Hoechst 33258染色检测细胞凋亡情况,并分别运用实时荧光定量PCR和Western blotting检测caspase 3在mRNA和蛋白水平的表达变化。结果Hoechst 33258细胞染色显示,对照组和SO处理组的细胞核为弥散均匀的圆形或椭圆形荧光,而ASO处理组的细胞核或细胞质内可见致密浓染的颗粒、新月体或环状荧光,核固缩,部分切片可见核碎裂。PCR和Western blotting检测显示,与对照组和SO处理相比,ASO处理组caspase 3 mRNA和蛋白的表达均明显上调。结论c-fos反义探针可以诱导人肝癌HepG2细胞凋亡,且caspase 3参与了此过程。
Objective To investigate the effect of c-fos antisense probe on the apoptosis of HepG2 cells and the role of caspase3 in it. Methods Human hepatoma HepG2 cells were divided into three groups: ① control group: 10μl normal saline; ② c-fos sense oligonucleotide (SO) group: SO10μl (5μg / μl); ③c-fos antisense oligonucleotide (ASO) treatment group: ASO 10 μl (5 μg / μl) was added. Each group of cells cultured for 1h after the follow-up experiments. The cell apoptosis was detected by Hoechst 33258 staining, and the expression of caspase 3 at mRNA and protein level was detected by real-time fluorescence quantitative PCR and Western blotting respectively. Results Hoechst 33258 staining showed that the nuclei of the control and SO treated groups were evenly distributed circular or oval fluorescence. In the nuclei or cytoplasm of ASO treated group, densely-stained particles, crescentic or circular fluorescence were observed, Nuclear pyknosis, some sections visible nuclear fragmentation. PCR and Western blotting showed that the expression of caspase 3 mRNA and protein in ASO group was significantly up-regulated compared with the control group and SO group. Conclusion The c-fos antisense probe can induce the apoptosis of HepG2 cells, and caspase 3 is involved in this process.