抗人DR5单克隆抗体对人肝癌细胞系HepG2的凋亡作用

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。

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

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 cytometry,and the cytotoxicity induced by mDRA-6 on HepG2 cells was assayed with MTT methodThe rate of apoptosis was determined by flow cytometry 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 mDRA6.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 cytometry assay.with Annexin V-FITC/PI staining.It is evident from the above observations that the anti-human DR5 monoclonal antibody can induce apoptosis of HepG2 cells.in vitro.