目的研究PEG10基因RNA干扰对肝癌细胞系Hep3B体外增殖的影响。方法psiRNA-PEG10真核表达载体转染Hep3B细胞,MTT法检测细胞体外增殖速度,流式细胞仪检测细胞周期变化。结果psiRNA-PEG10真核表达载体转染Hep3B细胞后,明显沉默了Hep3B mRNA和蛋白的表达,转染后48 h抑制效率最高。Hep3B细胞体外增殖受到抑制(P〈0.05),流式细胞仪分析结果显示大量Hep3B细胞凋亡,激光共聚焦显微镜观察到凋亡细胞形态。结论PEG10基因RNA干扰在体外可明显沉默肝癌细胞内PEG10基因的表达,抑制肝癌细胞的生长,诱导细胞凋亡。
Objective To investigate the effect of PEG10 gene RNAi (PEG10-RNAi) on proliferation of human hepatocellular carcinoma cell line Hep3B in vitro. Methods PEG10-RNAi expression vector was transfected to hepatocellular carcinoma cell line Hep3 B. Proliferation was assessed by MTT assay. Apoptosis of cells was evaluated by flow cytometry and confocal microscopy. Results Expressions of PEGIO mRNA and protein were remarkably reduced after transfection of PEGIO-RNAi for 48 h, and Hep3B cell growth was also inhibited by PEG10-RNAi significantly (P 〈 0.05). Apparent apoptosis was exhibited by flow cytometry analysis after PEG10-RNAi trasfected, and there was significant difference compared with control group (P 〈0.01 ). Apoptotic cells were revealed by confocal microscopy. Conclusion Hep3B cell proliferation was inhibited by PEG10-RNAi, and apoptosis was induced in vitro.