摘要
目的研究X-连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis,XIAP)siRNA或x1AP拮抗剂Embelin对肿瘤坏死因子相关凋亡诱导配体(tumor necros isfactor-relatedapoptosis—in—ducingligand,TRAIL)所致肝癌细胞HepG2生长抑制和细胞凋亡的影响。方法HepG2细胞转染xIAPsiRNA或者阴性对照,然后给予TRAIL处理。此外,HepG2细胞给予TRAIL、Embelin或者联合处理。XIAP表达水平变化、生长抑制、caspase-3活性分别用Westernblot、MTT测定、caspase-3荧光法检测。切割PARP的表达水平用Westernblot测定。结果XIAP蛋白表达水平在转染XIAPsiRNA后显著下调。与阴性对照相比,XIAPsiRNA显著增强TRAIL在100ng/ml(6.8%4-1.2%比11.8%±4.0%,P〈0.05)和1000ng/ml(18.9%±2.0%比26.6%±1.5%,P〈0.01)的生长抑制作用。在转染xIAPsiRNA后,TRAIL诱导caspase-3的活性和PARP的切割显著增强。此外,Embe—lin显著增强TRAIL对HepG2细胞的生长抑制(P〈0.01)、caspase-3活化(P〈0.01)和PAPR切割。结论XIAPsiRNA或Embelin在肝细胞癌的临床治疗方面具有潜在应用前景。
Objective To investigate the effects of x-(-linked inhibitor of apoptosis (XIAP) siRNA or XIAP antagonist Embelin on the growth inhibition and apoptosis of hepatcellular cancer cell line HepG2, which was treated by tumor necrosis factor-related apoptosis (TRAIL). Methods HepG2 cel[s were tranfected either by XIAP siRNA or a negative control, followed by treatment with TRAIL, Embelin or a combination of the two. XIAP expression, cell growth, and caspase-3 activity were determined by Western blot, MTT assay and fluorescent caspase-3 assay, respectively. Cleaved PARP expression levels of were measured by Western blot. Results The XIAP protein expression was significantly downregulated by transfection with XIAP siRNA. Compared with the negative con-trol, the XIAP siRNA significantly inhibited the growth of HepG2 cells treated by 100ng/ml(6.8%±1.2% vs. 11.8%±4.0%,P〈0.05)and1000 ng/ml (18.9%±2.0% vs. 26.6%±1.5%, P〈0.01) by TRAIL. TRAIL-induced caspase-3 activation and PARP cleavage were also increased significantly by XIAP siRNA transfection. In addition, Embelin also significantly inhibited cell growth (P〈0.01), activation of caspase-3 (P〈 0.01) and TRAIL-induced PARP cleavage. Conclusions Both XIAP siRNA and Embelin may potentially contribute to clinical treatment of hepatocellular carcinoma.
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2012年第5期381-385,共5页
Chinese Journal of Hepatobiliary Surgery