抗人DR5单克隆抗体对人肝癌细胞系HepG2的凋亡作用

Apoptosis of human hepatocellular carcinoma cell line HepG2 induced by anti-human DR5 monoclonal antibody

探讨抗人DR5单克隆抗体(mDRA-6)对人肝癌细胞系HepG2致凋亡的作用。常规培养人肝癌细胞系HepG2,流式细胞术定量分析检测HepG2细胞表面DR5的表达。MTT法检测mDRA-6的细胞毒性作用;Annexin V-FITC/PI双色标记HepG2细胞,流式细胞术定量分析检测细胞凋亡率;在荧光显微镜下观察mDRA-6对HepG2细胞形态变化的影响。结果显示,HepG2细胞表面有DR5表达,其平均表达百分率为40%。MTT法检测显示在40 mg/L mDRA-6浓度下可杀伤42%的细胞;Hoechst33258染色证实杀伤作用是通过细胞凋亡实现的经流式细胞术检测显示,3 mg/L的mDRA-6作用HepG2细胞6 h导致24.61%的细胞发生凋亡;在荧光显微镜下可观察到mDRA-6诱导导致HepG2细胞呈现典型细胞凋亡的形态特征。上述结果表明,mDRA-6能够诱导人肝癌细胞系HepG2凋亡。

To evaluate the effect of anti-human DR5 monoclonal antibody （mDRA-6） on the apoptosis of human hepatocellular carcinoma cell line HepG2, the surface expression of DR5 molecule on HepG2 cells was detected by flow eytometry, and the eytotoxieity indueed by mDRA-6 on HepG2 cells was assayed with MTT method The rate of apoptosis was determined by flow eytometry with Annexin V-FITC/PI staining, and the influence of mDRA-6 on the morphology of HepG2 cells was observed under fluorescent microscopy. In the experimental results, it was found that the DR5 molecules could be detected on the surface of HepG2 cells with a positive detection rate of 40%. As demonstrated by the MTT assay, the inhibitory rate on the proliferation of HepG2 cells accounted up to be 42% under treatment of 40 mg/L of mDRA-6. The rate of apoptosis of cells induced by 3 mg/L of mDRA-6 was 24.61% in the presence of 3 mg/L of mDRA-6 for 6 hours, as demostrated by the analysis with flow eytometry assay, with Annexin V-FITC/PI staining. It is evident from the above observations that the anti-human DR5 monoelonal antibody can induce apoptosis of HepG2 cells, in vitro.