STAT3诱骗寡核苷酸对肝癌细胞Hep G2的影响

The effect of STAT3 decoy ODNs on the growth and apoptosis of Hep G2 cells

目的观察应用诱骗寡核苷酸(decoy ODNs)靶向阻断信号转导子和转录激活子3(signal transducer and activator of tran-scription 3,STAT3)对肝癌细胞Hep G2的影响,初步探讨其中的分子机制。方法体外合成的STAT3 decoy ODNs在脂质体的介导下转染Hep G2细胞,荧光显微镜和流式细胞仪检测转染效率;通过细胞生长曲线观察对细胞增殖的影响;annexin-V/PI染色检测细胞凋亡;实时定量PCR检测STAT3下游基因bcl-xL、cyclin D1和c-myc的表达。结果荧光显微镜和流式细胞仪检测表明decoy ODNs能有效转染Hep G2细胞,转染效率达90.2%;生长曲线显示decoy ODNs有效抑制Hep G2细胞增殖;decoy ODNs处理细胞48 h后,流式细胞仪annexin-V/PI染色检测到早期细胞凋亡率为(22.86±2.63)%,晚期细胞凋亡率为(23.00±2.76)%;实时定量PCR证实decoy ODNs下调bcl-xL、cyclin D1和c-myc mRNA表达。结论STAT3 decoy ODNs能抑制细胞增殖,促进细胞凋亡,其作用机制可能与下调STAT3下游bcl-xL、cyclin D1和c-myc基因的表达相关。

Abstract：Objective To investigate the effect of STAT3 decoy ODNs on the growth and apoptosis of hepatoma cells Hep G2 and the related mechanism. Methods STAT3 decoy ODNs, synthesized in vitro, was transferred into Hep G2 cells by cationic lipid. The cell growth curve was used to reflect the growth and proliferation capacity of Hep G2, and annexin-V/PI staining method was used to calculate apoptic cells by FCM. Real time-PCR was used to quantitatively detect the expression of bcl-xL, cyclin D1, and c-myc mRNA in Hep G2. Results FCM assay and fluorescence microscopy demonstrated that STAT3 decoy ODNs was successfully transferred into Hep G2 cells （90.2%）. Decoy ODNs inhibted the growth of Hep G2 cells. After treated with decoy ODNs 48 h,the percentage of early apoptotic cells was （22.86 ± 2.63）% and that of late apoptotic cells was （23.00 ± 2.76）%. Real time-PCR indicated that Hep G2 cells transfected with STAT3 decoy ODNs had decreased the mRNA expression of bcl-xL,cyclin D1 and c-myc. Conclusions STAT3 decoy ODNs inhibited the growth of Hep G2 cells and induce the apoptosis. The mechanism may be associated with decreased expression of bcl-xL,cyclin DI and c-myc mRNA.