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探讨柚皮素增强抗人DR5单克隆抗体对人肝癌细胞系HepG2的凋亡作用及可能机制。MTT法检测柚皮素及抗人DR5单抗对人肝癌细胞系HepG2的生长抑制作用;Annexin V/PI双染分析细胞凋亡;蛋白免疫印记检测Caspase 8的表达;流式细胞术检测HepG2细胞表面DR5的表达。结果显示,10 mg/L的mDRA-6作用于HepG2细胞12 h后细胞增殖抑制率为39%;10 mg/L的mDRA-6分别与200μmol/L、400μmol/L、800μmol/L柚皮素共同作用于HepG2细胞后细胞增殖抑制率分别增加至58%、79%、85%;Annexin V/PI双染及流式细胞术检测结果表明,2 mg/L的mDRA-6作用细胞12 h后细胞凋亡率为16%,2 mg/L的mDRA-6和400μmol/L柚皮素共同作用于HepG2细胞12 h后细胞凋亡率增加为63%;mDRA-6和柚皮素共同作用于HepG2细胞后,蛋白免疫印记结果显示Caspase 8的激活片段明显显现,流式细胞术检测HepG2细胞表面DR5的表达,其基础表达率为49%,400μmol/L柚皮素作用于HepG2细胞12后,HepG2细胞表面DR5的表达率增加为81%。实验显示柚皮素可增强抗人DR5单克隆抗体对人肝癌细胞系HepG2的凋亡作用,其可能机制为柚皮素诱导HepG2细胞表面DR5受体表达上调所致。
To investigate the effect of naringenin on the apoptosis of human hepatoma cell line HepG2 induced by anti-human DR5 monoclonal antibody and its possible mechanism. The inhibitory effects of naringenin and anti-human DR5 McAb on the growth of human hepatocellular carcinoma cell line HepG2 were detected by MTT assay. The apoptosis of HepG2 cells was analyzed by Annexin V / PI double staining. The expression of Caspase 8 was detected by Western blotting. The expression of Caspase 8 was detected by flow cytometry Surface DR5 expression. The results showed that the inhibitory rate of mDRA-6 cells treated with 10 mg / L mDRA-6 for 12 h was 39%, while 10 mg / L mDRA-6 cells were incubated with 200 μmol / L, 400 μmol / L and 800 μmol / L naringenin After treated with 2 mg / L mDRA-6 for 12 h, the inhibitory rates of proliferation in HepG2 cells increased to 58%, 79% and 85%, respectively. Annexin V / PI double staining and flow cytometry showed that, The apoptosis rate was 16%. The apoptosis rate of HepG2 cells treated with 2 mg / L mDRA-6 and 400 μmol / L naringenin increased by 63%. The combination of mDRA-6 and naringenin HepG2 cells, Western blotting results showed that the active fragment of Caspase 8 was obvious, the expression of DR5 was detected by flow cytometry on the surface of HepG2 cells, the basic expression rate was 49%, 400μmol / L naringenin effect on HepG2 cells 12, The expression of DR5 on HepG2 cells increased by 81%. Experiments show that naringenin can enhance the anti-human DR5 monoclonal antibody on human hepatocellular carcinoma cell line HepG2 apoptosis, its possible mechanism for naringenin-induced HepG2 cell surface DR5 receptor expression due to the rise.