抗人DR5单链抗体诱导HepG2细胞凋亡的实验研究

目的探究抗人DR5单链抗体(scFv)ZF1对肝癌细胞株HepG-2的凋亡作用及机制。方法 MTT法检测ZF1对HepG-2的细胞毒效应;流式细胞术检测ZF1诱导HepG2的凋亡率;荧光显微镜观察经ANNEXIN-V/PI双染的HepG-2细胞;DNA Ladder检测HepG-2细胞的DNA特点;Western blotting检测Bcl-2、PARP蛋白的表达。结果 MTT结果显示ZF1抑制HepG-2细胞生长呈剂量依赖性,ZF1终浓度分别为0.225、0.45、0.9mg/ml、1.2mg/ml200μl时,HepG-2细胞的生长抑制率分别为28.8%、52.3%、65.3%、89.8%;流式细胞仪检测结果显示,终浓度分别为0.225、0.45、1.2mg/ml2ml作用HepG-2细胞4h,凋亡率分别为32.9%、56%、83.2%;荧光显微镜下可见早期凋亡细胞,细胞膜呈绿色荧光,细胞内有凋亡小体的形成,伴有核结构的变化;DNALadder出现明显的彗星尾状条带;Western blotting检测到ZF1诱导凋亡的细胞内Bcl-2、PARP蛋白表达上调。结论 ZF1可诱导HepG2细胞株凋亡,这种作...

Apoptosis of hepG2 cells induced by anti-human DR5 scFv(scFv) ZF1

We aimed to explore the apoptotic effects of anti-human DR5 scFv ZF1 on the hepatoma cell line HepG-2 and its mechanism.We employed MTT assay to detect the ZF1 cytotoxicity against HepG-2 cell,and found that ZF1 inhibited the growth of HepG-2 cells in a dose-dependent manner(ZF1 final concentrations of 0.225,0.45,0.9,1.2 mg/ml correspond to HepG-2 cell growth inhibition rates of 28.8%,52.3%,65.3%,89.8%);HepG2 cells were respectively treated with 0.225,0.45,1.2 mg/ml ZF1 for 4 hours,and flow cytometry indica...